【摘要】： 目的 建立人脂肪來源基質(zhì)細(xì)胞(hADSCs)的原代培養(yǎng)方法,并進(jìn)行脂肪誘導(dǎo)定向分化。探討凍存后的ADSCs的特點和脂肪分化潛能,為臨床應(yīng)用提供實驗室資料。 方法 取人腹部皮下脂肪,用酶消化法分離出單個核細(xì)胞,觀察其生長情況。細(xì)胞凍存于液氮中6個月,復(fù)蘇后觀察其生長變化和脂肪分化潛能。細(xì)胞脂肪誘導(dǎo)以含地塞米松、1-甲基-3-異丁基-黃嘌呤、胰島素的培養(yǎng)基作為脂肪定向誘導(dǎo)培養(yǎng)基。以油紅O染色檢測細(xì)胞內(nèi)脂質(zhì)沉積,觀察細(xì)胞形態(tài)學(xué)變化和計數(shù)分化細(xì)胞數(shù)。 結(jié)果 原代培養(yǎng)的細(xì)胞形態(tài)均一,呈成纖維樣,增殖力旺盛,可持續(xù)培養(yǎng)至15代。脂肪定向誘導(dǎo)培養(yǎng)后,第3天就可見細(xì)胞內(nèi)核周有脂肪滴沉積。細(xì)胞形態(tài)的變化和油紅O染色都證明這些細(xì)胞內(nèi)有脂肪的沉積。誘導(dǎo)第9天細(xì)胞內(nèi)出現(xiàn)大量脂質(zhì)沉積,在17天左右達(dá)到高峰。2周以后,大約90%左右的細(xì)胞定向分化為成熟的脂肪細(xì)胞。凍存和新鮮的ADSCs生長特點相似,脂肪分化潛能沒有差別(P0.5)。 結(jié)論 在成熟的脂肪組織中存在基質(zhì)細(xì)胞,具有良好的自我增殖和脂肪分化潛能。凍存后的細(xì)胞仍能繼續(xù)傳代培養(yǎng)。并且能保持與凍存前相似的的脂肪分化潛能。ADSCs可用于脂肪組織工程種子細(xì)胞來源。
[Abstract]:Objective to establish a primary culture method of human adipose derived stromal cells (hADSCs) and to induce adipose differentiation. To explore the characteristics and adipose differentiation potential of frozen ADSCs, and to provide laboratory data for clinical application. Methods Mononuclear cells were isolated from human abdominal subcutaneous fat by enzyme digestion, and the growth of mononuclear cells was observed. The cells were frozen in liquid nitrogen for 6 months and their growth and adipose differentiation potential were observed after resuscitation. The adipose induction medium contained dexamethasone, 1-methyl-3-isobutyl xanthine and insulin. Oil red O staining was used to detect lipid deposition, morphological changes and number of differentiated cells were observed. Results the cells cultured in primary culture were homogeneous in morphology, fibroblast-like, strong in proliferation, and could be continuously cultured for 15 generations. Fat droplets were observed around the nucleus of the cells on the third day after adipose induction. Changes in cell morphology and oil red O staining all demonstrated fat deposition in these cells. On the 9th day after induction, a large number of lipid deposits appeared in the cells, and reached the peak at about 17 days. After 2 weeks, about 90% of the cells differentiated into mature adipocytes. The growth characteristics of frozen ADSCs were similar to those of fresh ADSCs, and there was no difference in adipose differentiation potential (P0. 5). Conclusion there are stromal cells in mature adipose tissue with good self-proliferation and adipose differentiation potential. Cryopreserved cells can continue to be subcultured. ADSCs can be used as a seed cell source for adipose tissue engineering.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Raquel Taléns-Visconti;Ana Bonora;Ramiro Jover;Vicente Mirabet;Francisco Carbonell;José Vicente Castell;María José Gómez-Lechón;;Hepatogenic differentiation of human mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells[J];World Journal of Gastroenterology;2006年36期

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