Stem Cell Characterization

With Contract Assay Services

Stem cells are rare cells that may be phenotypically identified by the expression of cell surface markers. The identification of hematopoietic stem and progenitor cells (HSPCs) and mesenchymal stem cells (MSCs) is the result of years of scientific research showing that phenotypically-identified populations of cells maintain corresponding functions. Functional assays are the most stringent test of a stem cell’s identity and are key in understanding the effect of genes, biochemical pathways and candidate therapeutics.

Contract Assay Services can provide both phenotypic and functional assessments of your stem cell population of interest. Our scientists are experienced in designing and performing studies to evaluate the effects of novel test articles on the expansion and differentiation capacity of stem cells.

Contact us at contractassay@stemcell.com to learn how to take advantage of our stem cell characterization services and assays described below.

Hematopoietic Stem and Progenitor Cell Characterization

To measure the effects of test articles on hematopoietic stem and progenitor cell (HSPC) numbers and function, in vivo and in vitro assays have been developed. We can evaluate HSC renewal, expansion and ability to differentiate to various mature cell types. These assays include:

  • Engraftment assays
    Complex in vivo assay that allows the determination of the true hematopoietic stem cell (HSC) frequency and effects of test articles on this frequency by transplanting human stem cells into the NOD/SCID mouse model1

  • Long-Term Cultures - Initiating Cell (LTC-IC) assays
    Assay to determine the effect of test articles on the in vitro growth of primitive HSPCs using co-culture with stromal cells2
  • Mobilization assays
    Evaluation of the movement in vivo of HSCs out of bone marrow into peripheral blood following treatment with agents such as Mozobil and cytokines such as G-CSF (Figure 1)

  • Hematopoietic lineage-specific expansion cultures
    Ex vivo expansion of HSPCs (CD34+ cells) isolated from human cord blood or bone marrow and differentiation into erythroid, megakaryocyte or myeloid progenitor cells

  • Flow cytometry and profiling of various cell populations
    Including megakaryocytes, myeloid, erythroid, T, B and NK cells

Example Data from Previous Studies

Figure 1. Mobilization of Hematopoietic Progenitors Induced by rhG-CSF and AMD3100

C3H/HeN mice were treated with 5 µg rhG-CSF per day on days 0 - 2. Sixteen hours after the last G-CSF injection, one group of 5 mice was treated with 5 mg/mL of AMD3100 (also know as Mozobil or Plerixafor). Ninety minutes after treatment with AMD3100, mice were sacrificed and peripheral blood (PB) was collected. The progenitor content in the PB of each mouse was then assessed using the CFU assay. Mice treated with rhG-CSF alone showed a 10-fold increase in the number of CFUs per mL of PB, and mice treated with rhG-CSF and AMD3100 showed a 50-fold increase in the number of CFUs per mL of PB, when compared with vehicle control.

References

1. Szilvassy SJ et al. (2002) Quantitation of Murine and Human Hematopoietic Stem Cells by Limiting-Dilution Analysis in Competitively Repopulated Hosts. In: Klug CA & Jordan CT (Eds.), Methods in Molecular Medicine: Hematopoietic Stem Cell Protocols (pp. 167-187). Totowa, NJ: Humana Press.
2. Miller CL & Eaves CJ. (2002) Long-Term Culture-Initiating Cell Assays for Human and Murine Cultures. In: Klug CA & Jordon CT (Eds.), Methods in Molecular Medicine: Hematopoietic Stem Cell Protocols (pp. 123-141). Totowa, NJ: Humana Press.

Mesenchymal Stem Cell Characterization

Mesenchymal stem cells (MSCs) are a heterogeneous population of plastic-adherent, fibroblast-like cells, which are able to self-renew and differentiate into bone, adipose and cartilage tissue. In recent years there has been increased interest in MSCs and their potential utility in both tissue engineering and repair. MSCs also exhibit immunomodulatory and anti-proliferative effects on T cells, an attractive feature for cell therapy. Contract Assay Services can assess your MSC cell populations and provide assays for:

  • MSC expansion
    • Culture media that allows expansion using high quality media options including xeno- and animal component-free formulations

  • Characterization of surface markers
    • Including CD90, CD105, CD73, CD34 and CD45

  • In vitro differentiation into adipocytes, osteocytes and chondrocytes

  • Evaluation of the ability of MSC to suppress T cells (Figure 2)

Figure 2. MSC Suppression of T Cell Proliferation

Using co-culture experiments and the CFSE dye dilution assay, we can show that passage 2 MSCs can suppress division of activated T cells at both day 3 and day 7 of culture.

Custom Stem Cell Assays

STEMCELL Technologies is a world leader in providing products for stem cell research. We offer products and services for hematopoietic, mesenchymal and neural stem cell culture as well as embryonic stem (ES) cells and induced pluripotent stem (iPS) cells.

Get Started with Contract Assay Services Today

Our Assay Services Workflow

  1. One-on-one client consultation.
  2. Preparation of a proposal that clearly defines project scope, timeline and cost.
  3. Experimental execution and data analysis.
  4. Preparation of final report.

Contact STEMCELL Technologies

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