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
The type of assay you choose is dependent upon a number of factors, that include, but are not limited to:  
  • 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.
Contact an expert at Preferred Cell Systems™ at (719) 264-6251 or info@preferred-cell-systems.com for help and assistance.  

 
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