Megakaryopoietic Progenitor Cells
Megakaryocyte lineage proliferation is extremely limited. Once megakaryocytes are produced, their primary differentiation function is ploidy and the production of platelets.
Despite the low number of cell divisions within the megakaryopoietic lineage, Preferred Cell Systems™ has developed highly sensitive, instrument-based assays to selectively measure megakaryopoiesis in different tissues and species.
The traditional methylcellulose, colony-forming cell (CFC) assay for the detection of megakaryocyte colonies is rather insensitive. Nevertheless, megakaryocyte colonies derived from Mk-CFC can be detected using CFC assays from Preferred Cell Systems™.
Granulocyte-macrophage progenitor cells have traditionally been detected as colonies in methylcellulose colony-forming cell assays. These have been designated by different names, such as GM-CFC, CFU-GM, CFU, CFU-C. The colonies derived from these progenitors can be mixed, individual granulocyte (usually composed of neutrophils and designated G-CFC, or G-CFU) or macrophage colonies (M-CFC or M-CFU) and can be identified by their colony appearance.
Since the colony-forming assay cannot be used to measure cell proliferation, Preferred Cell Systems™ developed Suspension Expansion Culture™ (SEC™) Technology to provide an instrument-based measurement of Mk cell proliferation for several applications. Of importance, is the ability to combine these assays with other readouts, such for phenotypic identification by flow cytometry, thereby providing both proliferation and differentiation data using the same sample.
Preferred Cell Systems™ distinguishes between Mk progenitor cells cultured using SEC™ Technology and those grown in methylcellulose. All Mk progenitor cells using SEC™ technology are designated with the prefix "P" for "Progenitor".
In the table below, you will find the megakaryopoietic 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
- 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.
- Cell type being detected.
- Instrumentation available.
- Other factors such as accuracy, sensitivity, precision etc.
| SEC™ Mk Progenitor |
Use/ Application | SEC™ Assay / Reagent |
CFC Mk Cell |
CFC Assay | Growth Factor Cocktail | Primitiveness / Maturity |
|---|---|---|---|---|---|---|
| P-Mk 1 | Research or CellularTherapy |
CellExpand |
NA | NA | TPO,IL-3,SCF | Stem cell induction into primitive Mk compartment |
| P-Mk 1 | Research | CellExpand HALO HemoFLUOR HemoLIGHT |
Mk-CFC 1 | ColonyGro CAMEO-4 CAMEO-96 |
TPO,IL-3,SCF | Stem cell induction into primitive Mk compartment |
| P-Mk 1 | CellularTherapy | CellExpand HALO TE HemoFLUOR TE HemoLIGHT TE HALO PMT HemoFLUOR PMT HemoLIGHT PMT |
Mk-CFC 1 | ColonyGro CAMEO-4 CAMEO-96 |
TPO,IL-3,SCF | Stem cell induction into primitive Mk compartment |
| P-Mk 1 | In Vitro Toxicity | HALO-Tox HT | Mk-CFC 1 | ColonyGro CAMEO-4 CAMEO-96 |
TPO,IL-3,SCF | Stem cell induction into primitive Mk compartment |
| P-Mk 2 | NA | NA | Mk-CFC 2 | ColonyGro CAMEO-4 |
TPO alone | Mk compartment |
| All populations | Base growth medium for all uses | HemoGro |
Abbreviations and Comments
NA = Not available
Mk = Megakaryocyte
SEC™ = Suspension Expansion Culture™ Technology
P = Progenitor
Pr = Precursor
CFC = Colony-Forming Cell
IL-3 = Interleukin-3
TPO = Thrombopoietin
SCF = Stem Cell Factor