【摘要】： 目的:研究脂肪干細(xì)胞(adipose tissue-derived stem cells,ADSCs)在體外分離培養(yǎng)和在一定條件下誘導(dǎo)分化為脂肪細(xì)胞和成骨細(xì)胞的能力,探討其作為骨組織工程細(xì)胞的可行性。 方法:無菌切取SD大鼠腹股溝處的皮下脂肪,Ⅰ型膠原酶消化法獲取原代脂肪干細(xì)胞,接種至含10%胎牛血清的DMEM培養(yǎng)液,培養(yǎng)并適時(shí)傳代,每日在倒置顯微鏡下觀察并記錄細(xì)胞形態(tài)和增殖特征。第3代細(xì)胞分別在成脂成骨誘導(dǎo)液中誘導(dǎo)分化,并采用油紅染色,堿性磷酸酶(ALP)染色,Von Kossa染色進(jìn)行鑒定。 結(jié)果:從大鼠脂肪組織中分離出的脂肪干細(xì)胞,在體外呈成纖維細(xì)胞樣迅速增殖,可在異丁基-甲基黃嘌呤、地塞米松、胰島素等誘導(dǎo)下,表現(xiàn)出脂肪細(xì)胞特性;可以在地塞米松、抗壞血酸、β—甘油磷酸鈉的誘導(dǎo)下,表現(xiàn)出成骨細(xì)胞特性,該細(xì)胞堿性磷酸酶(ALP)染色ALP活性顯著增高,Von Kossa染色出現(xiàn)礦化結(jié)節(jié)。 結(jié)論:ADSCs易于在體外分離培養(yǎng),并擴(kuò)增迅速,其生物學(xué)特性穩(wěn)定,經(jīng)成脂誘導(dǎo)可分化為脂肪細(xì)胞,經(jīng)成骨誘導(dǎo)可分化為成骨細(xì)胞,可作為骨組織工程的種子細(xì)胞。
[Abstract]:Aim: to study the ability of adipose stem cells (adipose tissue-derived stem cells,ADSCs) to differentiate into adipocytes and osteoblasts in vitro and to explore its feasibility as bone tissue engineering cells. Methods: the subcutaneous fat of the groin of SD rats was cut aseptically and the primary adipose stem cells were obtained by type I collagenase digestion. The primary adipose stem cells were inoculated into the DMEM culture medium containing 10% fetal bovine serum. Cell morphology and proliferation were observed and recorded daily under inverted microscope. The third generation cells were induced to differentiate in adipogenic osteoblast and identified by oil red staining and alkaline phosphatase (ALP) staining with von Kossa staining. Results: adipose stem cells isolated from adipose tissue of rats proliferated rapidly as fibroblasts in vitro, and showed adipocyte characteristics induced by isobutyl-methylxanthine, dexamethasone, insulin and so on. The osteoblasts could be induced by dexamethasone, ascorbic acid and 尾 -glycerophosphate. The activity of ALP by alkaline phosphatase (ALP) staining was significantly higher than that by Von Kossa staining. Conclusion it is easy to isolate and culture in vitro and expand rapidly, and its biological characteristics are stable. The adipocytes can be differentiated into adipocytes by adipogenic induction and osteoblasts can be differentiated into osteoblasts by osteogenesis, which can be used as seed cells for bone tissue engineering.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 王燕,陳光輝,邵建華,田新利;大鼠脂肪組織源性間充質(zhì)干細(xì)胞的分離及向心肌細(xì)胞的誘導(dǎo)分化[J];山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2005年07期

2 鞠曉東,婁思權(quán),田華,王衛(wèi)國,劉延青;脂肪間充質(zhì)干細(xì)胞的基本生物學(xué)特性及向成骨細(xì)胞誘導(dǎo)分化的實(shí)驗(yàn)研究[J];中華實(shí)驗(yàn)外科雜志;2004年06期

3 雷永紅;付小兵;盛志勇;孫同柱;董曉紅;于勇;;誘導(dǎo)脂肪干細(xì)胞向表皮細(xì)胞表型的轉(zhuǎn)分化研究[J];中華實(shí)驗(yàn)外科雜志;2006年12期

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