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Hematopoietic Stem Cell and Progenitor Assays

ReachBio can perform hematopoietic stem cell and progenitor assays (i.e. CFC assays) using the cell types listed below. Some of these are human specific while others may be available from a number of species.

About Colony Forming Cell (CFC) Assays

FDA and/or other regulatory authorities are increasingly requesting that certain types of Colony Forming Cell (CFC) assays (in particular, CFU-GM for myelosuppression and BFU-E for anemia) be performed with drug candidates prior to Phase I clinical trials. Colony Forming Cell (CFC) assays are well characterized assays that can detect an increase or decrease in the frequency of hematopoietic progenitor proliferation in response to stimulatory or inhibitory (or toxic) agents. The relatively long incubation period (14 days) increases the sensitivity of the assay and allows the detection of more long-term effects than are detectable in a simple proliferation assay. Since cultures are assessed morphologically, distinct progenitor lineages can be discriminated and lineage-specific effects of compounds can be detected. 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. It is also possible to detect informative qualitative effects on individual cell morphology with CFC assays, even at compound concentrations that do not affect absolute colony number or proliferative potential. Furthermore, if desired, colonies can be individually removed from the semi-solid growth matrix for genotypic and/or phenotypic analysis of the (clonal) cell population. For more background information about CFC assays and their value in the drug development process, please click here.

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* ReachBio Study Directors are highly knowledgeable and have many years of experience in the scientific and practical aspects of CFC assays using a variety of cell sources from a variety of species.

Cell Samples Used by ReachBio in Colony Forming Cell (CFC) Assays

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CD34+ Cells

The CD34 cell surface marker is a glycoprotein found on a subset of human cells that correlate with hematopoietic stem and progenitor activity in vitro (via CFC assays) and in vivo (via engraftment). Substantial data supports the correlation of the number of non-expanded transplanted CD34+ cell numbers with clinical outcome, thus confirming the utility of this marker as a surrogate stem cell marker. ReachBio can perform hematopoietic CFC assays using CD34+ cells from a number of human cell sources, including:

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CD133+ Cells

The CD133 cell surface marker, also known as prominin, is found on a subset of CD34+ cells and is correlated with hematopoietic progenitor activity in vitro (via CFC assays). Although CD133+ cells have to date been less well studied than CD34+ cells in the clinical setting, evidence is accumulating that suggests that CD133 represents a useful surrogate stem cell marker. ReachBio can perform hematopoietic CFC assays using CD133+ cells from a number of human cell sources, including:

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Bone Marrow

Bone marrow is a rich source of both hematopoietic and non-hematopoietic stem and progenitor cells. It is therefore an excellent source of cells for in vitro CFC assays. ReachBio can perform CFC assays on bone marrow-derived cells, including:

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Cord Blood

Human cord blood is a rich source of hematopoietic and stem and progenitor cells. It is therefore an excellent source of cells for in vitro CFC assays. ReachBio can perform CFC assays on human cord blood-derived cells, including:

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Mobilized Peripheral Blood

Mobilized peripheral blood (MPB) is a rich source of hematopoietic stem and progenitor cells. It is therefore an excellent source of cells for in vitro CFC assays. ReachBio can perform CFC assays on mobilized peripheral blood-derived cells, including:

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Peripheral Blood

Compared to other cell sources such as mobilized peripheral blood, bone marrow and cord blood, peripheral blood is not a particularly rich source of hematopoietic stem and progenitor cells. However, cells with CFC activity are present within the blood at low frequency.  Due to the ease of access of this cell source and the potential for repeated samplings, it is often useful to perform CFC assays with blood-derived cells for certain applications, such as monitoring of CFC activity in patients participating in clinical trials. Other potential applications of CFC assays utilizing cells from peripheral blood include the detection of cytokines, chemokines or other agents with stem cell mobilizing activity. ReachBio can perform CFC assays on peripheral blood-derived cells, including: 

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Disease Specific Cells

Diseased cells often have altered receptor responses and/or signaling pathways. Therefore, responses of certain diseased cells to drug candidates may sometimes differ from those of their normal counterparts. Evaluation of drug candidates using primary human cells can be brought to an even greater level of relevance by utilizing qualified cells from patients with the specific diseases and comparing results to those obtained using normal cells. ReachBio can perform CFC assays using disease-specific cells. Availability and lead times may vary, depending upon the type of cell required.

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