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.
Gong
Wei
W
Department of Orthopaedics, XiJing Hospital, the Fourth Military University, Xi'an 710032, China; E-mail: gongwei@ fmmu. edu. cn.
Luo
Zhuo-Jing
ZJ
Han
Hua
H
Qin
Hong-Yan
HY
Chu
You-Biao
YB
Hu
Xue-Yu
XY
Lan
Li-Feng
LF