Isolation and culture of rat limbal stem cells and research on trans differation capacity
10.11659/jjssx.12E015112
- VernacularTitle:大鼠角膜缘干细胞分离培养及其横向分化能力初探
- Author:
Huiwen YAN
;
Jianying CHEN
;
Jie GAO
;
Wenping LIAO
;
Min SU
;
Rong HU
- Publication Type:Journal Article
- Keywords:
limbal stem cells;
neural stem cell-like cells;
epidermal growth factor;
basic fibroblast growth factor;
all-transretinoic acid
- From:
Journal of Regional Anatomy and Operative Surgery
2016;25(4):248-250,251
- CountryChina
- Language:Chinese
-
Abstract:
Objective To isolate and culture the rat limbal stem cells ( LSCs) in vitro,and investigate briefly their capacity for transdif-ferentiation into neural stem cells ( NSCs) with cytokine EGF,bFGF and RA.Methods LSCs derived from rats were cultured and identified by immunohistochemistry in vitro.LSCs were induced to differentiate into NSCs in the presence of EGF (20 ng/mL) ,bFGF (10 ng/mL) and with or without of RA(group 1 or group 2)(25 ng/mL) for seven days.Cultures without factors were used as control group.Then the neural marker Nestin of the coultured cells were measured by immunohistology staining.Furthermore,the positive cell rate was counted under micro-scope between the 2 groups and analyzed by statistical software.Results It showed that P63 was positive in LSCs.Nestin in both of the dif-ferentiation groups was positive at the rate of (77.01 ±6.32)%and (84.01 ±5.43)%respectively,of which the second group was higher than the first one (P<0.05).However,it was negative in the control group.A band of Nestin protein from cells was detected by western blot assay.Conclusion LSCs are successfully isolated and cultured in vitro.LSCs could be induced to differentiate into NSCs in the presence of EGF and bFGF.Moreover,the differentiation capability is enhanced in the condition of RA.
