大鼠脂肪干细胞向肠神经样干细胞转分化的初步实验研究  

A preliminary study on the transdifferentiation of adipose-derived stem cells into enteric nerve-like stem cells

作　　者：伍耀豪[1] 刘宝宁 陈露平 曾乐祥[1] 邱荣林[1] 张杰[1] 苏健航 廖敏仪 周军 邓小耿[1] WU Yao-hao;LIU Bao-ning;CHEN Lu-ping;ZENG Le-xiang;QIU Rong-lin;ZHANG Jie;SU Jian-hang;LIAO Min-yi;ZHOU Jun;DENG Xiao-geng(Department of Pediatric Surgery,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou,510120,China;Animal Center,School of medicine,Sun Yat sen University,Guangzhou 510080,China;Department of General Surgery,The Second Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830002,China)

机构地区：[1]中山大学孙逸仙纪念医院小儿外科,广州510120 [2]中山大学医学院动物中心,广州510080 [3]新疆医科大学第二附属医院普外科,新疆乌鲁木齐830002

出　　处：《岭南现代临床外科》2020年第2期146-150,共5页Lingnan Modern Clinics in Surgery

基　　金：广东省医学科研基金(A2018033);新疆自治区自然科学基金(2016D01C202)。

摘　　要：目的初步探讨应用小分子SB431542、noggin和LDN1931897将大鼠脂肪干细胞(ADSCs)转分化成肠神经样干细胞的可行性,为先天性巨结肠的细胞治疗提供一种新的种子细胞。方法取SD大鼠腹股沟脂肪组织分离培养ADSCs。当ADSCs传代至第3代时,加入小分子SB431542、noggin、LDN1931897以及肠神经培养基进行转分化实验。通过光学显微镜观察细胞形态学变化和应用免疫荧光染色检测其肠神经干细胞特异性标记物表达情况。结果ADSCs在含有3种小分子的肠神经细胞培养基中转分化培养7天后形成神经球样的细胞团。免疫荧光染色法检测发现神经干细胞特异性标记物Nestin呈阳性表达,肠神经干细胞标记物Sox2呈阳性表达。结论小分子SB431542、noggin和LDN1931897可在肠神经培养基中将ADSCs转分化为肠神经样干细胞。ADSCs有可能作为先天性巨结肠干细胞移植的种子细胞来源。Objective To explore the feasibility of using small molecules SB431542,noggin and LDN1931897 to transdifferentiate rat adipose-derived stem cells(ADSCs)into enteric neural-like stem cells,and to provide a new seed cell for cell therapy of congenital megacolon.Methods ADSCs were isolated and cultured from inguinal adipose tissue of SD rats.When ADSCs were passaged to the third generation,small molecules SB431542,noggin,LDN1931897,and enteric nerve medium were added for transdifferentiation experiment.The morphological changes of cells were observed by optical microscope,and the expression of enteric neural stem cells markers of was detected by immunofluorescence staining.Results ADSCs formed neurosphere-like cell clusters 7 days after transdifferentiation in enteric nerve cell culture medium containing three small molecules.Immunofluorescence staining showed positive expression of neural stem cell-specific marker Nestin and positive expression of intestinal neural stem cell marker Sox2.Conclusion Our preliminary experiments showed that small molecules SB431542,noggin,and LDN1931897 can transdifferentiate ADSCs into enteric neural-like stem cells in enteric nerve medium.ADSCs may serve as a seed cell source for congenital megacolonic stem cell transplantation.

关 键 词：脂肪干细胞 小分子 转分化 肠神经干细胞 

分 类 号：R656.9[医药卫生—外科学]