学术
干细胞体外自发分化和相关基因表达状况的研究
钱坤[1] 陈红[2] 章汉旺[1] 靳镭[1] 李豫峰[1] 刘群[1] 张苏明[2] 朱桂金[1]
[1]华中科技大学同济医学院附属同济医院生殖医学中心,武汉430030 [2]华中科技大学同济医学院附属同济医院生神经科,武汉430030
摘 要:
目的 在人胚胎干细胞（hESC）体外自发分化过程中，检测子宫内膜发育相关基因的表达以及子宫内膜样组织结构，子宫内膜样细胞的存在。方法 悬浮法培养类胚体14d，10％中性甲醛固定，常规石蜡包埋切片，取具有子宫内膜样组织结构的切片行雌激素受体（ER）免疫组织化学染色。hESC贴壁分化10d，免疫荧光双标检测雌激素受体／波形蛋白（角蛋白）的表达。hESC贴壁分化5d，RT-PCR方法检测子宫内膜发育相关基因：Wnt4、Wnt7a、Wnt5a、Hoxa10、Hoxa11、ER的表达。结果 hESC贴壁分化5d后可检测到Wnt4、Wnt7a、Wnt5a、Hoxa10、Hoxa11、ER的表达。hESC贴壁分化10d后，部分细胞同时表达ER／波形蛋白（角蛋白）。培养14d类胚体内存在子宫内膜样的腺体结构，部分腺体结构ER表达阳性。结论 hESC自发分化过程中，可能部分细胞具有自发分化为子宫内膜干细胞，子宫内膜细胞的倾向，但需对整个发育过程进行进一步鉴定。[著者文摘]
文章出处:
《解剖学报》-2007年38卷1期 -81-84页
分 类 号:
文献标识码:
A
文章编号:
0529-1356（2007）01-81
RESEARCH ON SPONTANEOUS DIFFERENTIATION OF STEM CELLS AND RELATED GENE EXPRESSION PROFILE
QIAN Kun , CHEN Hong , ZHANG Han-wang , JIN Lei , LI Yu-Feng , LIU Qun , ZHANG Su-ming , ZHU Gui-jin （ 1. Reproductive Medicine Center; 2. Neurology Department, Tongji Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China）
Abstract:
Objective To investigate the expression of endometrial developing-related genes, the endometrium-like structure and cells during spontaneous differentiation of hESCs. Methods （ 1 ） Embryoid bodies were cultured in suspension for 14days, fixed in 10% neutral-buffered formalin and embedded in paraffin. Estrogen receptor （ER） was detected by immunocytochemical staining. （2） hESCs were cultured in a 35 mm plastic dish and differentiated spontaneously for 10 days. The expression of ER and Vimentin/Keratin was detected by double immunolluorescence histochemistry. （3） hESCs were cultured in a 35 mm plastic dish and differentiated for 5 days. The expressions of endometrial developing-related genes including Wnt4, Wnt7a, Wnt5a, Hoxa10, Hoxa11 and ER were detected by RT-PCR. Results （1） The endometrium-like structures were detected in 14-day-old EBs and some were positive for ER staining. （2） Some spontaneously differentiated cells from hESCs for 10 days were positive for ER together with Vimentin/Keratin. （3）Wnt7a, Wnt4, Hoxa10, Hoxa11 and ER were detected by RT-PCR in hESCs which were differentiated spontaneously for 5 days. Conclusion During the spontaneous differentiation of hESCs, some cells are liable to differentiate into endometrial stem cells and endometrium-like cells. However, further identifications of the whole development process are needed to be done in the future.[著者文摘]
Key words:
Endometrium ; Endometrial stem cell; RT-PCR ; Double immunofluorescence histochemistry ;Differentiation; Human embryonic stem cell
基金资助:
国家自然科学基金资助项目（30370505）;同济医院基金资助
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