中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
27期
4988-4991
,共4页
唐欣%王岩%易海波%庞天舒
唐訢%王巖%易海波%龐天舒
당흔%왕암%역해파%방천서
干细胞%脐带脐血干细胞%脐带间充质干细胞%干细胞培养与分化%心肌细胞%基因表达%5-氮胞苷%反转录-聚合酶链反应%干细胞图片文章
榦細胞%臍帶臍血榦細胞%臍帶間充質榦細胞%榦細胞培養與分化%心肌細胞%基因錶達%5-氮胞苷%反轉錄-聚閤酶鏈反應%榦細胞圖片文章
간세포%제대제혈간세포%제대간충질간세포%간세포배양여분화%심기세포%기인표체%5-담포감%반전록-취합매련반응%간세포도편문장
stem cells%umbilical cord/umbilical cord blood stem cells%umbilical cord mesenchymal stem cells%stem cell culture and differentiation%cardiomyocytes%gene expression%5-azacytidine%reverse transcription-PCR%stem cel photographs-containing paper
背景：脐带来源的间充质干细胞进行自体心肌内移植，修复损伤心肌组织，是心血管研究领域的热点。目的：应用5-氮胞苷诱导人脐带间充质干细胞转化为心肌样细胞后进行鉴定，以确定心肌样细胞的结构、形态和功能。方法：分离培养人脐带间充质干细胞，在第2代细胞中分别加入浓度为2.5，5，10，20，40，80μmol/L 的5-氮胞苷，作用24 h 后除去，继续培养4周。结果与结论：5-氮胞苷诱导前，无细丝样结构、无颗粒，胞浆量均匀且少，核/浆比例高，细胞形态呈现典型的梭形，细胞呈漩祸样或同心圆生长，核内会见到较为明显的核仁；在5-氮胞苷处理24 h 之后，各组细胞都会出现部分死亡，失去典型的梭形形态，成为棍状或者柱状，40，80μmol/L 组细胞形态变化明显。反转录-聚合酶反应检测2.5，40μmol/L 组诱导4周，5，10，20μmol/L 组诱导1，2，3，4周时人脐带间充质干细胞心钠素、α-骨骼肌动蛋白基因表达呈阳性。提示经5-氮胞苷诱导的人脐带间充质干细胞表达心肌细胞的特定基因。
揹景：臍帶來源的間充質榦細胞進行自體心肌內移植，脩複損傷心肌組織，是心血管研究領域的熱點。目的：應用5-氮胞苷誘導人臍帶間充質榦細胞轉化為心肌樣細胞後進行鑒定，以確定心肌樣細胞的結構、形態和功能。方法：分離培養人臍帶間充質榦細胞，在第2代細胞中分彆加入濃度為2.5，5，10，20，40，80μmol/L 的5-氮胞苷，作用24 h 後除去，繼續培養4週。結果與結論：5-氮胞苷誘導前，無細絲樣結構、無顆粒，胞漿量均勻且少，覈/漿比例高，細胞形態呈現典型的梭形，細胞呈漩禍樣或同心圓生長，覈內會見到較為明顯的覈仁；在5-氮胞苷處理24 h 之後，各組細胞都會齣現部分死亡，失去典型的梭形形態，成為棍狀或者柱狀，40，80μmol/L 組細胞形態變化明顯。反轉錄-聚閤酶反應檢測2.5，40μmol/L 組誘導4週，5，10，20μmol/L 組誘導1，2，3，4週時人臍帶間充質榦細胞心鈉素、α-骨骼肌動蛋白基因錶達呈暘性。提示經5-氮胞苷誘導的人臍帶間充質榦細胞錶達心肌細胞的特定基因。
배경：제대래원적간충질간세포진행자체심기내이식，수복손상심기조직，시심혈관연구영역적열점。목적：응용5-담포감유도인제대간충질간세포전화위심기양세포후진행감정，이학정심기양세포적결구、형태화공능。방법：분리배양인제대간충질간세포，재제2대세포중분별가입농도위2.5，5，10，20，40，80μmol/L 적5-담포감，작용24 h 후제거，계속배양4주。결과여결론：5-담포감유도전，무세사양결구、무과립，포장량균균차소，핵/장비례고，세포형태정현전형적사형，세포정선화양혹동심원생장，핵내회견도교위명현적핵인；재5-담포감처리24 h 지후，각조세포도회출현부분사망，실거전형적사형형태，성위곤상혹자주상，40，80μmol/L 조세포형태변화명현。반전록-취합매반응검측2.5，40μmol/L 조유도4주，5，10，20μmol/L 조유도1，2，3，4주시인제대간충질간세포심납소、α-골격기동단백기인표체정양성。제시경5-담포감유도적인제대간충질간세포표체심기세포적특정기인。
BACKGROUND: Intramyocardial transplantation of autologous umbilical cord-derived mesenchymal stem cells for repair of myocardial tissue damage is paid increasing attention in the cardiovascular field. OBJECTIVE: Human umbilical cord mesenchymal stem cells were isolated and cultured. Passage 2 human umbilical cord mesenchymal stem cells were treated with various concentratins of 5-azacytidine (2.5, 5, 10, 20, 40, 80 μmol/L) for 24 hours . After removal of 5-azacytidine, cells were cultured for another 4 weeks. RESULTS AND CONCLUSION: Before 5-azacytidine treatment, filament-like structures or particles were not observed in the cells, but the amount of cytoplasm was less and uniform, nuclear/cytoplasm ratio was high, cells exhibited typical fusiform appearance and grew in a swirl-like manner, and nucleolus was obvious. After treatment with 5-azacytidine for 24 hours, some cells died in each group, and typical fusiform appearance turned into stick-like or column-like appearance, especial y in the 40 and 80 μmol/L 5-azacytidine groups. Reverse transcription-PCR results showed that atrial natriuretic peptide and α-skeletal actin gene expression levels were detected on human umbilical cord mesenchymal stem cells after treatment with 2.5 or 40 μmol/L 5-azacytidine for 4 weeks or with 5, 10, 20 μmol/L 5-azacytidine for 1, 2, 3 and 4 weeks. These findings suggest that 5-azacytidine-induced human umbilical cord mesenchymal stem cells express the specific gene of myocardiocytes.
