CAJ | 학술논문

目的观察复方麝香注射液在体外对人脐带间充质干细胞(hUMSCs)神经分化的影响.方法取足月妊娠剖宫产健康新牛儿脐带,获取间充质干细胞,采用流式细胞术进行细胞表型鉴定.以碱性成纤维细胞生长因子(bFGF)预诱导24 h,再以含5、10、15、20、25g/L复方麝香注射液的无血清培养基进行神经分化诱导,采用免疫细胞化学检测神经元特异性烯醇化酶(NSE)、微观相关蛋白-2(MAP-2)和胶质纤维酸性蛋白(GFAP)的表达.结果流式细胞检测显示hUMSCs表达CD13、CD29、CD44、CD73、CD90、CD105、CD106、CD166和HLA-ABC抗原,不表达CD31、CD34、CD38、CD45、CD133和HLA-DR抗原.经不同浓度复方麝香注射液诱导5 d后,各诱导组中均出现神经样细胞.5、10、15、20、25 g/L组显微镜每随机视野中NSE阳性细胞数分别为19.00±3.61、66.60±5.37、60.20±10.06、44.80±9.44、47.80±10.45,MAP-2阳性细胞数分别为9.20±3.27、53.40±10.92、59.40±10.41、46.60±8.88、46.80±8.93.以10 g/L和15 g/L组中的NSE、MAP-2阳性细胞数为最多.各诱导组中仅极少数细胞呈GAFP弱阳性.结论人脐带富含问充质干细胞(MSCs),适量复方麝香注射液可有效诱导人脐带间充质干细胞分化为神经元细胞.
목적 관찰복방사향주사액재체외대인제대간충질간세포(hUMSCs)신경분화적영향.방법 취족월임신부궁산건강신우인제대,획취간충질간세포,채용류식세포술진행세포표형감정.이감성성섬유세포생장인자(bFGF)예유도24 h,재이함5、10、15、20、25g/L복방사향주사액적무혈청배양기진행신경분화유도,채용면역세포화학검측신경원특이성희순화매(NSE)、미관상관단백-2(MAP-2)화효질섬유산성단백(GFAP)적표체.결과 류식세포검측현시hUMSCs표체CD13、CD29、CD44、CD73、CD90、CD105、CD106、CD166화HLA-ABC항원,불표체CD31、CD34、CD38、CD45、CD133화HLA-DR항원.경불동농도복방사향주사액유도5 d후,각유도조중균출현신경양세포.5、10、15、20、25 g/L조현미경매수궤시야중NSE양성세포수분별위19.00±3.61、66.60±5.37、60.20±10.06、44.80±9.44、47.80±10.45,MAP-2양성세포수분별위9.20±3.27、53.40±10.92、59.40±10.41、46.60±8.88、46.80±8.93.이10 g/L화15 g/L조중적NSE、MAP-2양성세포수위최다.각유도조중부겁소수세포정GAFP약양성.결론 인제대부함문충질간세포(MSCs),괄량복방사향주사액가유효유도인제대간충질간세포분화위신경원세포.
Objective To observe the effects of compound musk injection on neurocyte-like differentiation of human umbilical cord-derived mesenchymal stem cells (hUMSCs) in vitro. Methods The full-term umbilical cords from healthy neonates were washed with D-Hank' s,and MSCs were separated from them. After flow cytometry was performed,hUMSCs were pre-induced with 10 μg/L basic fibroblast growth factor (bFGF) ,then were induced to neural differentiation by serum-free culture medium containing different concentrations of compound musk injection including 5,10,15,20,25 g/L respectively. The expression of neuron specific enolase (NSE), microtubule-associated protein 2 (MAP-2) and glial fibrillary acid protein (GFAP) was detected by immunocytochemistcy. Results Fluorescence activating cell sorter (FACS) showed that hUMSCs expressed CD13, CD29, CD44, CD73, CD90, CD105, CD106, CD166, HLA-ABC (HLA-I) ,and did not express CD31 ,CD34,CD38,CD45,CD133,HLA-DR (HLA-Ⅱ). After induction for 5 days by DMEM/F12 medium containing different concentrations of compound musk injection, the hUMSCs of each group could differentiate into neuron-like cells. The number of NSE positive cells in 5,10,15,20,25 g/ L groups was 19.00 ±3.61,66.60 ±5.37,60.20 + 10.06,44.80 ±9.44,47. 80 ± 10.45,and that of MAP-2 positive cells was 9.20 ± 3. 27,53.40 ± 10. 92,59.40 ± 10. 41,46. 60 ± 8. 88,46. 80 ± 8. 93, respectively. NSE and MAP-2 positive cells were most plentiful in 10 g/L and 15 g/L groups. But just few of GFAP weak positive cells were detected in all five groups. Conclusion Human umbilical cord is a rich source of MSCs. Adequate concentration of compound musk injection can effectively induce the differentiation of hUMSCs into neural-like cells.