Hematopoietic Stem Cells
Preferred Cell Systems™ has developed tools to detect and measure up to 12 different stem cell populations depending on the application, media formulation, assay and species.
Hematopoiesis stem cells have usually been detected using the traditional methylcellulose, colony-forming cell (CFC) or colony-forming unit (CFU) assay.
In addition to offering CFC/CFU assays, Preferred Cell Systems™ also has in vitro stem cell expansion reagents and assays that do not incorporate methylcellulose. Instead, hematopoietic stem cells are grown in an advanced culture system called Suspension Expansion Culture™ (SEC™) Technology that not only allows cell-cell interaction, but also many other aspects to accurately and reliably detect stem cell characteristics.
Since one of the primary functions of stem cells is their potential and ability to proliferate, assays incorporating SEC™ Technology only use the most advanced and, in many cases, standardized and validated, instrument-based detection systems that allow even the rarest of stem cells to be detected. These assays have also been designed so that they can be combined with other readouts, such for phenotypic identification by flow cytometry and genetic analysis to obtain as much information as possible from a single sample.
Preferred Cell Systems™ distinguishes between hematopoietic stem cells grown in methylcellulose and those cultured using SEC™ Technology by their prefix nomenclature. Virtually all hematopoietic stem cells detected in methylcellulose will have the prefix "CFC". All hematopoietic stem cells using SEC™ technology with have the prefix "SC".
In the table below, you will find all the erythropoietic populations developed by Preferred Cell Systems™ that can be detected using either a SEC™-based or a methylcellulose assay. These are:
- CellExpand = Flexible readout of cell expansion
- HALO = Bioluminescence readout with standardization included
- HemoGLO™ = Bioluminescence readout with optional standardization
- HemoFLUOR = Fluorescence readout
- HemoLIGHT = Absorbance/colorimetric readout
- ColonyGro & CAMEO-4 = Manual or automated colony counting
- CAMEO-96 = Manual or automated colony counting following by bioluminescence measurement
- Use or application goal (research, cellular therapy or toxicity testing).
- Whether you need to measure proliferation, differentiation or both.
- Stem cell population being detected.
- Instrumentation available.
- Other factors such as accuracy, sensitivity, precision etc. Please note that rare stem cell populations cannot be detected using the CFC assay.
| SEC™ Stem Cell Population |
Use / Application | SEC™ Assay / Reagent |
CFC Stem Cell Population |
CFC Assay | Growth Factor Cocktail | Primitiveness / Maturity |
|---|---|---|---|---|---|---|
| Multipotent/ Multilineage |
Research/ CellularTherapy |
MultiCellGro |
Multipotent, Multilineage |
AllColonies | NA | Very primitive stem cell |
| SC-HPP X | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
NA | NA | IL-3, IL-6,SCF, Flt3-L | Quiescent L-H stem cell |
| SC-HPP 1 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
NA | NA | IL-3 ,IL-6, SCF, TPO, Flt3-L | Quiescent L-H stem cell |
| SC-HPP XT | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
NA | NA | IL-3,IL-6, SCF, Flt3-L, CD3, CD28 | Quiescent L stem cell |
| SC-HPP 2 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
NA | NA | EPO, GM-CSF, TPO, IL2 ,IL-3, IL-6, IL-7, SCF, Flt3-L | Induced primitive L-H stem cell |
| SC-HPP 2 | Cellular Therapy | HALO QC HALO SC-IPS HALO PMT HemoGLO PMT HemoFLUOR PMT HemoLIGHT PMT |
NA | NA | EPO, GM-CSF, TPO, IL2 ,IL-3, IL-6, IL-7, SCF, Flt3-L | Induced primitive L-H stem cell |
| SC-HPP 2 | In Vitro Toxicity | HALO-Tox HT | NA | NA | EPO, GM-CSF, TPO, IL2 ,IL-3, IL-6, IL-7, SCF, Flt3-L | Induced primitive L-H stem cell |
| SC-HPP 1T | Research | HALO HemoGLO HemoFLUOR HemoLIGHT |
NA | NA | IL-3, IL-6, SCF, Flt3-L, IL-2, CD3, CD28 | Induced primitive L stem cell |
| SC-GEMM 1 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
CFC-GEMM 1 "MethoCult Express" |
ColonyGro CAMEO-4 CAMEO-96 |
EPO, GM, IL-3, IL-6, SCF, TPO, Flt3-L | Induced primitive H stem cell |
| SC-GEMM 1 | Cellular Therapy | CellExpand STEMpredict HALO QC HALO SC-IPS HALO PMT HemoGLO PMT HemoFLUOR PMT HemoLIGHT PMT HALO-Tox HT |
CFC-GEMM 1 "MethoCult Express" |
ColonyGro CAMEO-4 CAMEO-96 |
EPO, GM, IL-3, IL-6, SCF, TPO, Flt3-L | Induced primitive H stem cell |
| SC-GEMM 1 | In Vitro Toxicity | HALO-Tox HT | CFC-GEMM 1 "MethoCult Express" |
ColonyGro CAMEO-4 CAMEO-96 |
EPO, GM, IL-3, IL-6, SCF, TPO, Flt3-L | Induced primitive H stem cell |
| SC-GEMM 2 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
CFC-GEMM 2 | ColonyGro CAMEO-4 CAMEO-96 |
EPO, GM, IL-3, IL-6, SCF, TPO | Induced H stem cell |
| SC-GEMM 3 | Research/ Cellular Therapy |
CellExpand HemoGLO PCA HemoFLUOR PCA HemoLIGHT PCA |
CFC-GEMM 2 (MethoCult H4435 "Enriched") |
ColonyGro CAMEO-4 |
EPO, GM, IL-3, IL-6, SCF, Flt3-L TPO | Induced H stem cell |
| SC-GEM 1 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
CFC-GEM 1 | ColonyGro CAMEO-4 CAMEO-96 |
EPO, GM, IL-3, IL-6, SCF | Mature H stem cell |
| SC-GEM 2 | Research | CellExpand HALO HemoGLO HemoFLUOR HemoLIGHT |
CFC-GEM 2 (MethoCult H4434 "Classic") |
ColonyGro CAMEO-4 |
EPO, GM, IL-3, SCF | Mature H stem cell |
| SC-GEM 2 | Cellular Therapy | CellExpand HemoGLO PCA HemoFLUOR PCA HemoLIGHT PCA |
CFC-GEM 2 (MethoCult H4434 "Classic") |
ColonyGro CAMEO-4 |
EPO, GM, IL-3, SCF | Mature H stem cell |
| SC-GEM 3 | Cellular Therapy | HemoGLO PCA HemoFLUOR PCA HemoLIGHT PCA |
CFC-GEM 3 | ColonyGro CAMEO-4 |
EPO,GM,G,IL-3,SCF | Most mature H stem cell |
| All populations | Base growth medium | HemoGro |
Comments
Note: " TPO" = TPO added CFC-GEMM 3, but not present in equivalent competitor reagent.
CFC-GEMM can be used to detect colonies from all hematopoietic lineages including progenitor and precursor cells.
Abbreviations
SC = Stem Cell
L-H = Lympho-Hematopoietic
L = Lymphopoietic
H = Hematopoietic
CFC = Colony-Forming Cell
HPP = High Proliferative Potential
GEMM = Granulocyte, Erythroid, Macrophage, Megakaryocyte
GEM = Granulocyte, Erythroid, Macrophage
IL-2 = Interleukin 2
IL-3 = Interleukin-3
IL-6 = Interleukin 6
EPO = Erythropoietin
TPO = Thrombopoietin
GM = Granulocyte/Macrophage Colony Stimulating Factor
G = Granulocyte Colony Stimulating Factor
SCF = Stem Cell Factor
Flt3-L = Flt3-Ligand