本文選題：成人脂肪干細胞 + 體外誘導(dǎo)��； 參考：《山東醫(yī)藥》2017年40期

【摘要】：目的觀察成人脂肪干細胞(ADSCs)的體外多向分化能力。方法從正常成人脂肪組織分離得到ADSCs并體外培養(yǎng)。在ADSCs中分別加入成人脂肪、骨、軟骨、肝細胞的體外誘導(dǎo)培養(yǎng)基誘導(dǎo),觀察誘導(dǎo)過程中細胞形態(tài)的變化,誘導(dǎo)前、誘導(dǎo)1周、誘導(dǎo)2周時采用RT-PCR法觀察細胞相應(yīng)標(biāo)志物mRNA表達的變化。結(jié)果 ADSCs向脂肪細胞誘導(dǎo)2周,細胞內(nèi)的脂質(zhì)空泡聚集,小脂滴融合在一起并變大;最后細胞變?yōu)閱畏?并且充滿脂質(zhì),紅油O染色陽性;誘導(dǎo)1周細胞PPARγ、脂蛋白酶和αP2 mRNA表達較誘導(dǎo)前增加;誘導(dǎo)2周細胞PPARγ、脂蛋白酶mRNA表達降低,但αP2 mRNA變化不明顯。ADSCs向骨細胞誘導(dǎo)5天,細胞由成纖維形轉(zhuǎn)化為多邊形或不規(guī)則狀。誘導(dǎo)2周單層細胞聚集成塊,形成細胞島狀結(jié)構(gòu),ALP染色陽性,von Kossa染色可見黑色礦化結(jié)節(jié);礦化結(jié)節(jié)形成后細胞向結(jié)節(jié)移行聚集,促使更多的細胞形成更大的礦化結(jié)節(jié);誘導(dǎo)后細胞ALP、骨橋蛋白和Ⅰ型膠原mRNA表達逐漸增加。ADSCs向軟骨細胞誘導(dǎo)1周,細胞呈多邊形或類圓形,細胞團向內(nèi)呈放射狀聚集,FITC染色后免疫熒光可見Ⅱ型膠原;誘導(dǎo)1、2周時細胞Ⅱ型膠原、聚集蛋白聚糖和SOX9 mRNA表達明顯高于誘導(dǎo)前。ADSCs向肝細胞誘導(dǎo)2周時可見部分細胞變成圓形或橢圓形,并隨時間順延呈肝細胞樣圓形細胞增多趨勢,細胞核居中,圓形且邊緣清楚,PAS染色陽性;誘導(dǎo)1、2周時細胞CK18、HNF1α及白蛋白mRNA表達明顯高于誘導(dǎo)前。結(jié)論成人脂肪組織中分離得到的成人ADSCs可在體外定向分化為脂肪細胞、骨細胞、成骨細胞和肝細胞。
[Abstract]:Objective to observe the differentiation ability of adult adipose stem cells (ADSCs) in vitro. Methods ADSCs was isolated from normal adult adipose tissue and cultured in vitro. Adult fat, bone, cartilage and hepatocytes were added to ADSCs to induce in vitro. The changes of cell morphology were observed during the induction. The expression of mRNA was observed by RT-PCR method at 2 weeks after induction. Results after ADSCs was induced to adipocytes for 2 weeks, the lipid vacuoles gathered and the small lipid droplets fused together and became larger. Finally, the cells became monocyclic and filled with lipid, and the cells were positive for marked oil O staining. The expression of PPAR 緯, lipoprotein enzyme and 偽 P2 mRNA was increased at 1 week after induction, but the expression of PPAR 緯 and lipase mRNA decreased at 2 weeks after induction, but the change of 偽 P2 mRNA was not obvious. After 5 days of induction, the cells transformed from fibroblast to polygon or irregular. After 2 weeks, monolayer cells were induced to assemble into blocks to form the island structure of the cells, and black mineralized nodules could be seen by von Kossa staining, and after the formation of the nodules, the cells migrated to the nodules, which caused more cells to form larger mineralized nodules. After induction, the expression of ALP, osteopontin and type I collagen mRNA increased gradually. ADSCs were induced to chondrocytes for 1 week. The cells were polygonal or round. The expression of type 鈪，

本文編號：1919427