Isolation and differentiation of embryonic stem cells from BALB/c mouse.
Objective To investigate the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.
Department of Orthopaedics, XiJing Hospital, the Fourth Military University, Xi'an 710032, China; E-mail: gongwei@ fmmu. edu. cn.
Wei
W
Gong
Zhuo-Jing
ZJ
Luo
Hua
H
Han
Hong-Yan
HY
Qin
You-Biao
YB
Chu
Xue-Yu
XY
Hu
Li-Feng
LF
Lan