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ReachBio Research Labs

Seattle, Washington, US

ReachBio Research Labs provides primary cell biology based contract research services and products to the drug development and life science research communities.

With expertise encompassing many aspects of primary cell biology and a special focus on hematopoietic (blood and bone marrow) primary cell systems, ReachBio Research Labs works with customers and clients involved in basic research through multiple areas of pre-clinical and clinical drug development.

Our platform is broad and includes predictive and investigative toxicology, immunology, immuno-oncology, cell bank development, antibody testing, flow cytometry/cell sorting, assay development and much more.

ReachBio Research Labs has not listed any services.

Clinical Biomarkers
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Cytokine Storm Testing
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Our cytokine storm assay, which can be customized to meet the demands of your program, utilizes primary human PBMCs or NHP cells, sourced fresh (less than 6 hours old) to ensure maximum viability and function. The cells are incubated under positive control (anti-CD3, clone OKT3), carrier control (PBS or other) and test conditions... Show more »

Our cytokine storm assay, which can be customized to meet the demands of your program, utilizes primary human PBMCs or NHP cells, sourced fresh (less than 6 hours old) to ensure maximum viability and function. The cells are incubated under positive control (anti-CD3, clone OKT3), carrier control (PBS or other) and test conditions for 24 – 72 hours. The supernatants can then be assessed for pro-inflammatory cytokines using Luminex™ technology, and the cells themselves assessed for proliferation and activation markers by flow cytometry.

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ADCC and CDC Assays
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An important mechanism utilized by the cell-mediated immune system to target and kill virus-infected or other diseased cells (e.g. cancer / tumor cells) is through antibody-dependent cell-mediated cytotoxicity (ADCC). This mechanism has been adopted to create in-vitro cell-killing assays to ascertain the efficacy of therapeutic... Show more »

An important mechanism utilized by the cell-mediated immune system to target and kill virus-infected or other diseased cells (e.g. cancer / tumor cells) is through antibody-dependent cell-mediated cytotoxicity (ADCC). This mechanism has been adopted to create in-vitro cell-killing assays to ascertain the efficacy of therapeutic monoclonal antibodies (mAb) to kill targeted tumor cells. This assay relies on a process using three key components; an effector cell that is typically an NK cell, mAb (IgG class) and tumor cell. ADCC is initiated when the bifunctional mAb (typically IgG1) links to the effector cell and tumor cell creating a ‘bridge’ that allows for cell killing. This process triggers release of lytic proteins (perforins and proteases) from the effector cell that enter the tumor cell and kills it through lytic mediated apoptosis.

Peripheral blood mononuclear cells (PBMCs) as well as isolated Natural Killer (NK) cells can be used in ADCC assays. However, within PBMCs, the NK cell is the mediator of the ADCC process as the principal effector cell. In drug development, the ability of NK cells to bind monoclonal antibodies to target tumor cell killing is considered to be important to the efficacy of the therapeutic monoclonal antibody screened in ADCC assays.

While the Fab fraction (mAb) binds to targeted cell-surface antigens (of tumor), the Fc component binds to NK cell CD16 Fc-gammaIIIa (FCGR3A) receptors. Various clinical oncology studies with therapeutic mAb’s (e.g. rituximab targeting CD20, trastuzumab targeting HER2 and cetuximab targeting EGFR) have revealed observations that indicate enhanced NK cell killing activity of tumor cells in certain types of patients. Supporting studies show that people with a specific polymorphism for FCGR3A (Homozygous for V158+ CD16), display enhanced NK cell activity vs. other genotypes.

At ReachBio, we recognize the interest in utilizing pre-screened donor NK cells and incorporate them in our ADCC assays. For more information on these primary cells, please visit our product page.

Our ADCC assays allow clients to evaluate and rank their mAb candidates ability to kill tumor/cancer cells through NK mediated cell lysis.

Our ADCC Assay services:
• Allow clients’ to evaluate and rank their mAb(s) candidates effectiveness at killing tumor/cancer cells
• Are suitable for antibody drug research and development
• Are compatible with bi-specific antibodies and other immune cell recruiters
• Utilize Primary cells (PBMCs, unscreened NK cells or pre-screened NK V158+ cells) to enable more
clinically relevant decision-making
• Provide flow cytometry based readouts with additional intracellular analysis, if desired
• Can be customized to your program’s needs

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Immuno-oncology Assays
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Our customized, primary cell approach creates a test system that is as close as possible to the in vivo situation while allowing for robust data generation. Our immuno-oncology assays include autologous MLRs, mixed donor MLRs and clonogenic assays. Autologous MLRs allow us to evaluate the ability of a lead therapeutic to reverse... Show more »

Our customized, primary cell approach creates a test system that is as close as possible to the in vivo situation while allowing for robust data generation. Our immuno-oncology assays include autologous MLRs, mixed donor MLRs and clonogenic assays. Autologous MLRs allow us to evaluate the ability of a lead therapeutic to reverse T cell exhaustion and increase tumor cell killing directly in a donor sample. Mixed donor MLRs allow for the evaluation of checkpoint inhibitors (e.g. Tim-3), and cytokine release in a carefully controlled experimental model in which primary human effector cells (T cells or NK cells) are added at predetermined ratios. Checkpoint inhibitor assays are assessed by flow cytometry and cytokine release measured by Luminex™ . Clonogenic assays allow for the evaluation of the killing of the cancer stem/progenitor cell in an in vitro model (e.g. CFC assay).

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In vitro Hematoxicity Testing
Price on request

Neutropenia Assessment

Neutropenia is a condition where there are abnormally low numbers of neutrophils (a type of white blood cell) in the blood. Neutropenia is often a side effect of drug treatments and this effect may be acute or chronic and can become a rate limiting factor for continued treatment. Since neutrophils... Show more »

Neutropenia Assessment

Neutropenia is a condition where there are abnormally low numbers of neutrophils (a type of white blood cell) in the blood. Neutropenia is often a side effect of drug treatments and this effect may be acute or chronic and can become a rate limiting factor for continued treatment. Since neutrophils have very short half-lives, any drugs that inadvertently affect them can cause serious medical complications. Our scientists have the ability to evaluate the effect of test compounds on the neutrophil population through validated CFC (colony forming cell) assays and liquid culture assays. These assays may be performed on bone marrow derived from multiple species, including all animals which are used routinely in toxicology studies.

Assays:

Neutrophils are derived from precursor cells in the bone marrow. These cells undergo differentiation and proliferation to generate mature neutrophils. By adding the test compounds to bone marrow cells in a semi-solid methylcellulose matrix supplemented with cytokines (e.g. ColonyGEL™), we can evaluate their effect on the clonal growth (development of 3D colonies) of the CFU-GM (colony forming unit- granulocyte, macrophage) derived cell. IC50 values for compounds generated with this assay have correlated with clinical neutropenia. The CFU-GM assay has been validated by ECVAM and endorsed by the FDA. The myeloid liquid culture assay allows for the assessment of neutrophil generation, through analysis of antigen acquisition or loss associated with maturation. By adding the drugs to this culture system with primitive bone marrow cells and specialized cytokines, we can assess maturation by cytospin and flow cytometry.

Thrombocytopenia Assessment

Thrombocytopenia is a condition where there are abnormally low numbers of platelets in the blood. A normal platelet count may range from 150 – 450 x 106 per mL of blood. The function of platelets is to stop bleeding by clumping and clotting blood vessel injuries. Platelets are derived from the megakaryocytes within the bone marrow, therefore, if a drug treatment inadvertently targets the megakaryocytes, there will be a decreased production of platelets. Alternatively, a drug may inadvertently cause an increased destruction of the mature platelets, both scenarios will result in thrombocytopenia. We have two distinct assay platforms for thrombocytopenia that allow us to assess whether test compounds have an effect on the production or the destruction of platelets. These assays may be performed on bone marrow derived from multiple species, including many species that are used routinely in toxicology studies.

Assays:

The precursor cells go through proliferation, differentiation, and endoreduplication to generate platelets. By adding the test compounds to bone marrow cells in a collagen-based semi-solid matrix supplemented with cytokines, we can evaluate their effect on the clonal growth (development of 3D colonies) of the CFU-Mk (colony forming unit- megakaryocyte) derived cell. IC50 values generated with this assay can assess if compounds have an effect on platelet production. This in vitro assay has been prevalidated with support from ECVAM (European Centre for the Validation of Alternative Methods). Additionally, we have a megakaryocyte liquid culture system utilizing primitive bone marrow cells and specialized cytokines, which allows assessment of the maturing megakaryocytes and platelets by flow cytometry (CD41 expression).

Severe Anemia Assessment

Severe anemia is a condition where there are abnormally low numbers of red blood cells which may be the result of blood loss, an inability to produce adequate red blood cells or a breakdown of the red blood cell. Blood loss may result from injury, whereas the inability to produce red blood cells may be due to deficiencies in iron or vitamin B12. Another potential reason for a decrease in red blood cells may be side effects of certain drug treatments. Our scientists have the ability to evaluate the effect of test compounds on the red blood cell populations through CFC (colony-forming cell) assays and liquid culture assays. These assays may be performed on bone marrow derived from multiple species including many animals which are used routinely in toxicology studies.

Assays:

Red blood cells are derived from precursor cells in the bone marrow. These cells undergo differentiation and proliferation to generate reticulocytes and then mature red blood cells. By adding the test compounds to bone marrow cells in a semi-solid methylcellulose matrix supplemented with cytokines (e.g. ColonyGEL™), we can evaluate their effect on the clonal growth (development of 3D colonies) of the BFU-E (burst forming unit - erythroid) and CFU-E (colony forming unit - erythroid) derived cell with IC50 values recorded. This assay has been useful at predicting severe anemia clinically. By adding drugs to our erythrocyte liquid culture assay (containing primitive bone marrow cells and specialized cytokines), we can assess the maturation red blood cells, including reticulocytes (thiazole orange staining) and mature red blood cells by analysis of antigen acquisition or loss associated with maturation via flow cytometry.

Lymphopenia Assessment

Lymphopenia, or lymphocytopenia is the condition of having an abnormally low level of lymphocytes in the blood and may be caused by a reduction of T, B or NK cells or a combination of all three. Certain therapies, often (but not always) targeting NHL (Non-Hodgkin Lymphoma) related diseases, cause off target cell death of lymphocytes (T cells, B cells and NK cells). The toxicity can be species-dependent. For example, some drugs can cause severe lymphopenia in small animal models, but have little toxicity on human cells. This issue can make preclinical development challenging and frustrating. Many groups use small animal models in order to assess for lymphopenia risk. A finding of lymphotoxicity in one of these models may derail a program. Our customizable lymphotoxicity assays allows for the side-by-side assessment of lymphotoxicity risk in human, NHP, rat, mouse and dog lymphocytes, providing highly detailed information about interspecies differences in toxicities as well as a realistic evaluation of potential clinical toxicity.

Assays:

We collect fresh, primary blood from humans, mice, rats, dogs or NHPs under appropriate IRB or IACUC approval and then prepare the PBMCs (peripheral blood mononuclear cells). Our cytotoxic assays use PBMCs cultured in the presence of test articles and control compounds for 16 to 48 hours, following which the T, B and NK cells are assessed for necrosis and/or apoptosis by quantitative flow cytometry. The broad spectrum of available antibodies with multiple fluorochromes allows for the simultaneous analysis of the multiple lymphoid subpopulations within the same sample. Additionally, in order to assess if the compounds are acting on lymphoid precursors, CFC (colony-forming cell) assays can be employed. There is an excellent and robust Pre-B CFC assay using mouse cells, but lymphoid precursor assays for other species are limited.

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Antibody Testing
Price on request

We’ve worked with a large variety of client therapeutic antibody constructs and have established primary cell assays for ADCs, BiTE’s and immuno-oncology related antibody therapies that include human, NHP, mouse, rat and dog cell-based models.
We begin any antibody-related project by first designing a customized experimental... Show more »

We’ve worked with a large variety of client therapeutic antibody constructs and have established primary cell assays for ADCs, BiTE’s and immuno-oncology related antibody therapies that include human, NHP, mouse, rat and dog cell-based models.
We begin any antibody-related project by first designing a customized experimental layout that includes dosing, replicates, controls, cell type and source, and timing, along with any other important considerations. After a protocol is agreed upon, we source appropriate (e.g. endotoxin-free) reagents for both cell stimulation and experimental readouts. In order to obtain data that is as translatable as possible to the clinic, whenever appropriate we use the same antibodies and flow cytometry gating strategies that are used clinically in the diagnosis of hematological diseases.

Assay formats include:

Antibody Efficacy Testing
Antibodies may be evaluated on primary cells, cell lines and also on specific primary leukemia cells showing functionality and efficacy. These assays may involve co-culture of NK cells with primary cancer cells (e.g. MM or AML).
ADCC Assays
Primary human NK cells genotyped at the FCGRIII locus have been pre-screened by our scientists for use in these assays.
Redirected T Cell Killing
These assays use primary cells and cancer cell lines in co-culture systems (e.g. using HLA matched primary human PBMC’s or T cells).
T Cell Cytotoxicity
These assays can be performed with pan-T cells (CD3+) or T cell subsets (CD4+ or CD8+).
MLR Assays
Allogeneic mixed leukocyte reaction assays may involve T cells such as pan-T cells or T cell subsets with in vitro derived mDCs (monocyte-derived dendritic cells).

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Stem Cell Mobilization Assays
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Combined in vivo and in vitro assays for evaluating mobilization

Hematopoietic stem cell mobilizers are agents that cause hematopoietic stem cells to leave their resident niches within the bone marrow and transiently move (or “mobilize”) into the blood stream. The kinetics of mobilization can vary greatly, depending upon... Show more »

Combined in vivo and in vitro assays for evaluating mobilization

Hematopoietic stem cell mobilizers are agents that cause hematopoietic stem cells to leave their resident niches within the bone marrow and transiently move (or “mobilize”) into the blood stream. The kinetics of mobilization can vary greatly, depending upon the mobilizing agent and the dose. Other cell types may also be mobilized into the blood stream by the mobilizing agent. There can also be species-dependent differences in responses to the mobilizing agent which can be assessed at ReachBio Research Labs.
ReachBio Research Labs has worked with clients with both small and large molecule mobilizers in both rodent and NHP models. We have also analyzed mobilization efficacy from human patient samples. We can provide the in vivo dosing component for rodent (mouse) models, or receive mobilized rodent, NHP or human blood samples from the client or their third party in vivo service provider.

Typical Services for Assessing Stem Cell Mobilizing Agents Include:

• Standard blood cell counts
• Analysis of CD34+ (or mouse equivalent) cell content and other cell phenotypes via flow cytometry
• Evaluation of progenitor content by in vitro Colony Forming Cell (CFC) assays
• Evaluation of functional stem cell content by in vivo NOD/SCID reconstitution assays

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In vitro Radiosensitivity Assays
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Radioprotection and Radiorecovery Assays

Ionizing radiation (e.g. due to intensive radiotherapy, radiological/nuclear incident or act of terrorism) can produce deleterious dose-dependent effects in living tissues, leading to significant morbidity and a potentially fatal illness affecting various organs.
Radiosensitivity is... Show more »

Radioprotection and Radiorecovery Assays

Ionizing radiation (e.g. due to intensive radiotherapy, radiological/nuclear incident or act of terrorism) can produce deleterious dose-dependent effects in living tissues, leading to significant morbidity and a potentially fatal illness affecting various organs.
Radiosensitivity is the relative susceptibility of cells, tissues, organs, organisms, or other substances to the injurious effects of radiation. In general, it has been found that cell radiosensitivity is directly proportional to the rate of cell division and inversely proportional to the degree of cell differentiation. In short, this means that actively dividing cells or those not fully mature are most at risk from radiation. Therefore, there is great interest in the development of effective radioprotectors and radiorecovery drugs for the hematopoietic system, which is continuously producing cells.

Our Services:

Utilize primary hematopoietic progenitor and mesenchymal (stromal) cell assays to evaluate compounds for potential radioprotective and radiation-sparing effects

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Immunophenotyping
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Immune monitoring is an important part of many preclinical and clinical programs. T, B and NK cells can be enumerated along with more recently discovered cells such as MDSC (myeloid derived suppressor cells) and dendritic cells. In diseased patients, the presence of important markers (e.g. CD138 in multiple myeloma) can be... Show more »

Immune monitoring is an important part of many preclinical and clinical programs. T, B and NK cells can be enumerated along with more recently discovered cells such as MDSC (myeloid derived suppressor cells) and dendritic cells. In diseased patients, the presence of important markers (e.g. CD138 in multiple myeloma) can be monitored over the course of a clinical trial to assess if the treatment is having an effect on the diseased cells. We have the ability to measure up to 13 colors in a single sample and can do direct blood subset counts using our flow cytometer. We also have the ability to perform internal staining procedures. Immune monitoring may be performed on whole blood, PBMCs, and bone marrow and from a variety of species including human, NHP, mouse, rat and dog, as well as for a variety of disease types in humans including AML, MM, CML, PV, NHL, Sickle Cell Anemia, SLE and RA.

Assays:

Our Flow cytometry assays, using our BeckmanCoulter cytoFLEX™, allow multiple panels to be assessed for the relative frequency of a given cell population within a sample, assessment of activation markers and internal staining (e.g. foxP3 and pStat5) for measurement of phosphoproteins (e.g. TKI potency) and nuclear factors. Our readouts allow for numerous parameters to be assessed within biological samples that include cell enumeration and cell viability, in addition to cell subset analysis, cell apoptosis and cell necrosis.

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Drug Target Validation
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Target validation is the process by which newly discovered cellular pathways that are believed to play a role in diseases are confirmed. An important part of drug target validation is confirming that the therapeutic target is both present in the primary diseased cells, and that disruption of the target elicits a desired effect... Show more »

Target validation is the process by which newly discovered cellular pathways that are believed to play a role in diseases are confirmed. An important part of drug target validation is confirming that the therapeutic target is both present in the primary diseased cells, and that disruption of the target elicits a desired effect (e.g. induction of apoptosis). We use primary diseased cells, cultured under conditions that mimic the in vivo conditions as closely as possible, thus providing researchers with detailed, relevant information about the effects of their compounds. Our years of experience performing experiments on blood and bone marrow allow us to design experiments where cytokine release, cellular response and genomic data can be obtained from a single well of a 96-well tissue culture dish. Our focus on primary blood and bone marrow assays allows us to help researchers functionally validate targets in most of the important blood cancers (e.g. AML, CML, MM, NHL) and immune diseases.

Assays:

Our customized approach to target validation enables our clients to obtain large amounts of functional and genomic data from individual experiments. Cultured primary cells are exposed to test or control articles under appropriate conditions, and then a combination of colony forming cell assays (CFC), quantitative flow cytometry, cytokine measurement and genomic analysis are performed. This approach allows for a large amount of information to be obtained from samples that are often highly restricted in cell number.
Primary blood cancer cells may be cultured in both liquid medium or methylcelluose-based (e.g. ColonyGEL™) media for various target readouts by CFC assays. Following liquid culture, cells can be assessed for receptor quantitation. The CFC assays allow investigation of the effects of test compounds on blast and progenitor cancer cells, thus confirming activity of a test compound to a specific target. Flow cytometry is optimized to detect a novel target if that target is on the outside of the cell. Quantitation of this target can be used to assess if novel molecules are capable of upregulating or downregulating target expression. Receptor quantitation may also be performed. Clonogenic CFC assays may be used to assess compound specific activity against a target on a specific progenitor. Gene expression may be used to detect a novel molecular target.

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Cancer Stem Cell Tumor Models
Price on request

Hematopoietic Cancer Stem Cells and Leukemic Progenitor Cells

For various hematological diseases (e.g. chronic myeloid leukemia, acute myeloid leukemia, multiple myeloma, lymphomas), the molecular abnormalities and perturbed signaling pathways that give rise to these diseases have, in recent years, been elucidated.

In... Show more »

Hematopoietic Cancer Stem Cells and Leukemic Progenitor Cells

For various hematological diseases (e.g. chronic myeloid leukemia, acute myeloid leukemia, multiple myeloma, lymphomas), the molecular abnormalities and perturbed signaling pathways that give rise to these diseases have, in recent years, been elucidated.

In many, if not all forms of cancer, a small number of slowly dividing, self-renewing cancer stem cells give rise to abundant and rapidly dividing diseased cells. Targeted killing of the cancer stem cells, while retaining the normal hematopoietic progenitors, is a new focus of research, aiming to develop more effective and less toxic cancer treatments. We have the ability to monitor cancer stem cells in patients over the course of a clinical trial using in vitro assays with cells from patients. We can also assess the differential toxicity of compounds on cancer stem cells (leukemic colony forming cells) compared to normal hematopoietic progenitor cells.

Our Assay Services Include:

Common applications include prediction of compound and targeted cell killing for assays below

AML (acute myeloid leukemia) Progenitor Assays
• Evaluate effects of single agents or combined therapies
• Genotyping and other molecular analyses

ALL (acute lymphoblastic leukemia) Progenitor Assays
• Evaluate effects of single agents or combined therapies

CML (chronic myelogenous leukemia) Progenitor Assays
• Evaluate effects of single agents or combined therapies
• Genotyping (bcr-abl)

CLL (chronic lymphocytic leukemia) Progenitor Assays
• Evaluate effects of single agents or combined therapies

MM (multiple myeloma) Progenitor Assays
• Evaluate effects of single agents or combined therapies

NHL, MCL, DLBCL (lymphoma) Progenitor Assays
• Evaluate effects of single agents or combined therapies

PV (polycythemia vera) Progenitor Assays
• Evaluate effects of single agents or combined therapies
• Genotyping (jak2v617f)

AA (aplastic anemia)
• Combined Therapies

MDS (myelodysplastic syndromes)
• Combined Therapies

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Colony Formation Assays
Price on request

CFC (Colony-Forming Cell) Assays

CFC assays are well characterized and validated in vitro assays that can detect an increase or decrease in the frequency of hematopoietic progenitor proliferation and/or changes in differentiation potential in response to stimulatory, inhibitory or toxic agents. This is in sharp contrast to... Show more »

CFC (Colony-Forming Cell) Assays

CFC assays are well characterized and validated in vitro assays that can detect an increase or decrease in the frequency of hematopoietic progenitor proliferation and/or changes in differentiation potential in response to stimulatory, inhibitory or toxic agents. This is in sharp contrast to simple proliferation assays, which only provide an aggregate readout of all proliferating cells and cell types within a bulk culture. The FDA and other regulatory authorities are increasingly requesting these types of assays be performed with drug candidates prior to Phase I clinical trials in order to assess the potential hemotoxic liability of the compounds. Since cultures are assessed morphologically by highly trained microscopists, distinct progenitor lineages (myeloid, erythroid and megakaryocytic) can each be discriminated and lineage-specific effects of compounds can be detected. For example, decreases in myeloid colonies (CFU-GM) correlate with clinical neutropenia. Decreases in erythroid colonies (CFU-E and BFU-E) correlate with clinical anemia and decreases in megakaryocyte colonies (CFU-Mk) correlate with thrombocytopenia. If desired, colonies can be individually removed from the semi-solid growth matrix for genomic and/or phenotypic analysis of the (clonal) cell population.

Colony Assays are Predictive and Validated. The CFC assay has been found to be particularly valuable at predicting drug-induced neutropenia and myelosuppression and determining maximum tolerated dose (MTD). The CFU-GM assay has been validated by the European Commission for Alternative Methods (ECVAM). The CFU-Mk assay has been prevalidated with support from ECVAM. Multi-drug chemical entities were evaluated that include immunosuppressive compounds, anti-neoplastics, anti-virals and pesticides.

Types of Colony-Forming Cell Assays Performed by ReachBio Research Labs:

CFU-GM (Myeloid Progenitor Assays)
CFU-Mk (Megakaryocytic Progenitor Assays)
BFU-E (Erythroid Progenitor Assays)
CFU-Pre-B (Lymphoid Progenitor Assays)
CFU-F (Mesenchymal Progenitor Assays)
CFU-L (Leukemic Colony-Forming Cell Assays)

Additionally, we can perform CFC assays and other hematopoietic stem and progenitor cell assays using cells from multiple species.

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In vivo Assays
Price on request

NOD/SCID Repopulation Assays

The NOD/SCID mouse system is a powerful tool to assess true stem cell number and functional behavior. Although in vitro assays such as Colony Forming Cell (CFC) assays are extremely useful in enumerating and analyzing hematopoietic and non-hematopoietic progenitors, the only test for true stem... Show more »

NOD/SCID Repopulation Assays

The NOD/SCID mouse system is a powerful tool to assess true stem cell number and functional behavior. Although in vitro assays such as Colony Forming Cell (CFC) assays are extremely useful in enumerating and analyzing hematopoietic and non-hematopoietic progenitors, the only test for true stem cells is their ability to reconstitute the entire blood forming system of an ablated host. The NOD/SCID mouse is an immunodeficient recipient, which allows the introduction of human, NHP or mouse cells and the determination of stem cell functionality through engraftment, proliferation and differentiation into at least two distinct lineages (typically myeloid and lymphoid). This in vivo reconstitution assay is typically known as the Competitive Repopulating Unit (CRU) or SCID Repopulating Cell (SRC) assay.

Stem Cell Mobilization Assays

Hematopoietic stem cell mobilizers are agents that cause hematopoietic stem cells to leave their resident niches within the bone marrow and transiently move (or “mobilize”) into the blood stream. The kinetics of mobilization can vary greatly, depending upon the mobilizing agent and the dose. Other cell types may also be mobilized into the blood stream by the mobilizing agent. There can also be species-dependent differences in responses to the mobilizing agent which can be assessed at ReachBio Research Labs.

ReachBio Research Labs has worked with clients with both small and large molecule mobilizers in both rodent and NHP models. We have also analyzed mobilization efficacy from human patient samples. We can provide the in vivo dosing component for rodent (mouse) models, or receive mobilized rodent, NHP or human blood samples from the client or their third party in vivo service provider.

5-FU Assays

Transplantation & Engraftment Kinetics in 5-FU Model

The 5-FU model is an excellent and well documented in vivo assay for monitoring myelosuppression (or protection) of various agents.

A common side effect following chemotherapeutic agent administration is myelosuppression leading to neutropenia. 5-fluorouracil (5-FU) is a DNA replication inhibitor that kills rapidly dividing cells (including neutrophil progenitors) while sparing more quiescent cells (such as resting stem cells). The single dose administration of 5-FU to mice at 150 mg/kg causes a significant and almost immediate (within 2 days) reduction in the white blood cell, neutrophil, and femoral bone marrow cellularity. Over the next 12 days, there is an initial gradual recovery of the bone marrow progenitors followed by recovery of neutrophil cell numbers. Compounds that augment the hematopoietic (e.g. G-CSF) system can be evaluated with this model.

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Flow Cytometry
Price on request

Immune Profiling & Flow Cytometry Services

Biotherapeutics development often requires a deep understanding of their effects on the entire immune cell system. Our expertise includes flow cytometry profiling of cell surface markers and intracellular staining to look at master regulatory proteins. In addition to looking at... Show more »

Immune Profiling & Flow Cytometry Services

Biotherapeutics development often requires a deep understanding of their effects on the entire immune cell system. Our expertise includes flow cytometry profiling of cell surface markers and intracellular staining to look at master regulatory proteins. In addition to looking at T cells, B cells and NK cells, we can also examine helper T cell subsets (Th0, Th1, Th2, Th17 and Tregs) by both surface staining and intracellular staining.
Our experience also includes studying intracellular cytokine production in Th subsets by flow cytometry as well as quantitative measurements of secreted cytokines in primary cell cultures by ELISA or Luminex™.

ReachBio Research Labs Flow Cytometry Services Include:

Flow Cytometry
• Cell surface phenotyping based on surface marker staining
• Intracellular flow cytometry for intracellular cytokine, phosphoprotein or nuclear factor analysis
• We can perform up to 16 color flow cytometry
Cell Sorting
• Our cell sorting capabilities include 8 color gating combined with 4 channel sorting
Molecular Readouts
• Gene expression analysis by qPCR
• Multiplexed gene expression analysis
Cytokine Readouts
• Over 200 cytokines, chemokines and other growth factors are available for analysis in cell culture
supernatants

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Biochemistry & Molecular Biology
Price on request

Stand-Alone or Add-On Molecular Biology Services

Molecular biology gives a deep understanding of the molecular components of the cell, namely DNA, RNA and proteins. Our molecular biology experts work one-on-one with clients to determine a molecular approach that will help address questions from their program. For this... Show more »

Stand-Alone or Add-On Molecular Biology Services

Molecular biology gives a deep understanding of the molecular components of the cell, namely DNA, RNA and proteins. Our molecular biology experts work one-on-one with clients to determine a molecular approach that will help address questions from their program. For this reason, we design our molecular experiments based on client requirements, with common applications including isolating nucleic acids from blood and bone marrow and gene expression analysis on colonies from our primary
cell-based assay. Our molecular biology services are performed stand-alone or are paired with any of our in vitro, primary cell biology assays; the most popular being the myeloid, erythroid, and megakaryocyte assays.
We offer a range of services to accommodate both simple and complex requests. Don’t hesitate to inquire about how our molecular services can aid in moving your program forward.

Examples of Molecular Biology Services we offer, but are not limited to:

Genotyping
HLA
FCGR3A
BCR-ABL
Nucleic Acid
Isolation from (frozen or fresh) whole blood, and bone marrow
Isolation from any of our in vitro assays
Gene Expression
qPCR
Miscellaneous Services
Collect cells pellets or lysates from primary cell-based assays

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Cell Banking
Price on request

Culture expansion and banking of both primary cells and cell lines

If you have the need for high quality and reproducible replicate vials of cells for your drug development programs, we can help. We offer cell banking services for both human and mouse primary cells and cell lines. We have experience in banking even the... Show more »

Culture expansion and banking of both primary cells and cell lines

If you have the need for high quality and reproducible replicate vials of cells for your drug development programs, we can help. We offer cell banking services for both human and mouse primary cells and cell lines. We have experience in banking even the most challenging types of cells, including primary lymphocyte subsets requiring multiple rounds of stimulation-induced expansions prior to banking. For those requiring specialized pre-qualified NK cells for ADCC assays, we have genotyped multiple donors (V158+) and can bank these cells in customized sizes to support your research.

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Stable Cell Line Generation
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LTC-IC Assay
Long-Term Culture Initiating Cell Assay
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Long Term Bone Marrow Cultures were first described by Dexter et al. These co-culture systems allow for the continuous culture of bone marrow hematopoietic cells on an adherent layer of bone marrow stromal cells. Within this in vitro culture system, hematopoietic stem cells retain their “stemness” in that they continuously... Show more »

Long Term Bone Marrow Cultures were first described by Dexter et al. These co-culture systems allow for the continuous culture of bone marrow hematopoietic cells on an adherent layer of bone marrow stromal cells. Within this in vitro culture system, hematopoietic stem cells retain their “stemness” in that they continuously generate the various progenitors and mature progeny of multiple blood cell lineages and also can reconstitute the ablated hematopoietic system of irradiated mice. The progenitor cells generated in this system can be quantified and evaluated using colony-forming cell assays.

Typical Applications:

The Long Term Bone Marrow Culture system allows the continuous administration of test compounds and kinetic evaluation of their impact on hematopoietic stem cells and progenitor cells.
Evaluation of the ability of normal and diseased marrow stroma to support hematopoietic stem cell function

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Oncology Animal Models
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Clinical Chemistry
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Toxicology
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Bioanalysis
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Cell-Based Assays
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Drug Discovery
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Biomarkers
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Pharmacology & Toxicology
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Drug Discovery & Development
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Cytotoxicity Assays
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Functional & Cell Type Specific Assays
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Cell and Tissue Culture
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Protein Services
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Antibody Services
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Biospecimen Management and Storage
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Antibody/Antigen Detection Based Testing
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Animal Models and Studies
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Animal Physiology Analyses
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Lead Identification and Validation
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Specialized Cell-Based Assays
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Animal Models of Disease
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Biology
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Clinical Research
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Clinical Laboratory Services
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In vitro Toxicity Testing
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Cell Viability & Proliferation Assays
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Immunoassays
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Cells and Tissues
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Cytometry
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Biospecimens
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Animal Model in vivo Analyses
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2018-06-15 17:54:13 -0400

Net Promoter Score of 10 received for In vitro Hematoxicity Testing.

Additional Ratings: satisfaction with deliverable: 9, satisfaction with timeliness: 9.

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