摘要
目的 :观察胚胎肝Sca - 1+细胞能否在体外向神经组织细胞分化。方法 :免疫磁珠法分离孕 14 5dC5 7BL/6J小鼠的胚胎肝的Sca - 1+细胞 ,以DMEM/F12 + 10 %胎牛血清培养液培养 ,当细胞融合达 80 % ,按 1∶3比例传代。第 5代细胞用BME和RA先后诱导 2 4h ,再用无血清培养基培养 5h至 5d。免疫细胞化学检测细胞特点。结果 :胚胎肝Sca - 1+细胞经诱导后表现神经元形态 ,并表达神经元特异蛋白如神经元特异性核蛋白 (NeuN)、神经丝蛋白M、神经元特异性微管蛋白 1(TuJ- 1) ,但是无微管相关蛋白Tau、MAP - 2和星形胶质细胞特异酸性蛋白 (GFAP)表达。结论 :胚胎肝Sca- 1+细胞 (主要为造血干细胞 )具有可塑性 ,在体外能分化为早期未成熟的神经元样细胞。
AIM: To study whether Sca-1 + cells from fetal liver can be induced to differentiate into neuronal cells in vitro. METHODS: Sca-1 + cells from 14 5-days-old murine fetal liver were isolated with a magnetic cell sorting kit, and were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 supplemented with 10% fetal bovine serum (FBS), and passaged at a ratio of 1∶3 when cells reached more than 80% confluence. The 5 passage cells were induced by 10 -3 mol/L β-mercaptoethanol (β-ME) and 5 ×10 -7 mol/L all-trans-retinoic acid (RA) for 24 hours, and then incubated in serum-free medium for 5 hours to 5 days. The characteristics of treated cells were assayed by immunocytochemistry staining analysis at 5 hours, or 5 days.RESULTS: Cells treated with β-ME and RA exhibited neuronal phenotype and expressed neuron-specific protein such as neuron-specific nuclear protein (NeuN), neuronfilament-M, and neuron-specific tubulin-1 (TuJ-1) but not tau, MAP-2, or the astrocyte-specific marker glial fibrillary acidic protein (GFAP).CONCLUSION: Sca-1 + cells from fetal liver, of which most are regarded as hematopoietic stem cells, could differentiate into early immature neuronal cells in vitro . These findings suggest that Sca-1 + cells from fetal liver may be an alternative source in cell therapy and gene therapy of neural dysfunction.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2003年第9期1178-1181,T002,共5页
Chinese Journal of Pathophysiology
基金
SupportedbyScienceandTechnologyFoundationofGuangdongProvince (No .99M01204G),ScienceandTechnologyFoundationofGuangzhoucity (No .2 0 01-2-037-01),andKeyScienceFoundationofGuangdongProvince (No .0 2 1195 )
