A Novel Approach for Amplification and Purification of Mouse Oligodendrocyte Progenitor Cells.
Although transgenic and knockout mice are widely used to study the specification and differentiation of oligodendrocyte precursor cells (OPCs), mouse primary OPCs are difficult to be purified and maintained, and many in vitro studies have to resort to rat OPCs as substitutes. In this study, we reported that mouse O4 negative early-stage OPCs can be obtained by culturing cortical tissue blocks, and the simultaneous treatment of OPCs with Platelet Derived Growth Factor-AA (PDGFaa), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF) is the key for the propagation of mouse OPCs in culture. EGF was found to be a potent mitogen for OPCs and cooperate with PDGFaa to extend cell division and inhibit their differentiation. EGF also collaborates with PDGFaa and bFGF to convert bipolar or tripolar OPCs to more vital fibroblast-like OPCs without compromising their oligodendrocyte differentiation potential. In addition, EGF promoted the survival and proliferation of glial progenitor cells (GPCs) derived from primary OPC cultures, and a mixture of GPCs and OPCs can be obtained and propagated in the presence of EGF, bFGF, and PDGFaa. Once EGF is withdrawn, GPC population decreased sharply and fibroblast-like OPCs changed into typical OPCs morphology, then homogeneous OPCs were obtained subsequently.
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Junlin
J
Yang
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Xuejun
X
Cheng
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Jiaxi
J
Shen
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Binghua
B
Xie
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Xiaofeng
X
Zhao
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal University Hangzhou, China.
Zunyi
Z
Zhang
The Vivian L Smith Department of Neurosurgery, University of Texas Medical School at Houston, Houston TX, USA.
Qilin
Q
Cao
Department of Neurobiology, Key Laboratory of Medical Neurobiology of the Ministry of Health, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University School of Medicine Hangzhou, China.
Ying
Y
Shen
Zhejiang Key Laboratory of Organ Development and Regeneration, The Institute of Developmental and Regenerative Biology, College of Life and Environment Sciences, Hangzhou Normal UniversityHangzhou, China; Department of Anatomical Sciences and Neurobiology, University of Louisville, LouisvilleKY, USA.
Mengsheng
M
Qiu
EGF; GPCs; OPCs; PDGFaa; coculture; stratified culture; synergistic effect