Rat bone marrow derived mesenchymal progenitor cells support mouse ES cell growth and germ-like cell differentiation.
Mouse embryonic fibroblasts (MEFs) have been used as feeder cells to support the growth of mouse embryonic stem cell (mESC) and primordial germ cells (PGC) in culture for many years. However, MEF preparation is a complex and tedious task. Recently, there are reports indicating that the microenvironment provided by bone marrow stromal cells could support the survival of embryonic-like stem cells in bone marrow. In this report, rat bone marrow derived mesenchymal progenitor cells (MPC) were used as feeder cells to culture mouse Oct4-GFP ES cell and ES cell derived germ cells. FACS results show that similar to MEF, rat MPC could efficiently support growth of the mouse Oct4-GFP ES cell line in culture (MPC 85.5 +/- 5.1% vs MEF 84.1 +/- 6.2%). ES cells could be subcultured for >15 passages without losing morphological characteristics. The cultured cells expressed stem cell marker alkaline phosphatase, Oct4, Sox2, and SSEA-1. Furthermore, rat MPC cells were able to support survival of germ cells isolated from mouse Oct4-GFP ES cell formed embryoid bodies (EB). After induction by retinoic acid for 7 days, some isolated cells differentiated to spermatogonial stem-like cells, expressing Mvh, Stra-8, Hsp90-a, integrinb1 and a6. Compared with traditional MEF culture systems, the rat MPC culture system is effective in supporting ES cell growth and is easy to prepare.
Cui
Guanghui
G
Medical Center of Peking University & Hong Kong Science and Technology University, Guangdong, China.
Qi
Zhengyu
Z
Guo
Xin
X
Qin
Jie
J
Gui
Yaoting
Y
Cai
Zhiming
Z
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D005854:Germ Cells / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism
D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins