【摘要】：【目的】對(duì)兔脂肪來(lái)源的干細(xì)胞（rabbitadipose-derivedstemcells，RADSCs）的分離、培養(yǎng)和鑒定進(jìn)行探討，并初步探討不同濃度轉(zhuǎn)化生長(zhǎng)因子β3（TransformingGrowthFactor-β3，TGF-β3）促進(jìn)RADSCs向軟骨細(xì)胞分化的可行性。為軟骨組織工程中種子細(xì)胞的選擇提供新思路。 【方法】取成年大耳白兔腹股溝部或頸背部脂肪組織，利用酶消化法和密度梯度離心法獲得細(xì)胞；進(jìn)行原代和傳代培養(yǎng)，，觀察細(xì)胞變化并繪制細(xì)胞生長(zhǎng)曲線；流式細(xì)胞術(shù)和免疫熒光技術(shù)鑒定細(xì)胞表面CD分子；將獲取的第三代細(xì)胞進(jìn)行體外成軟骨細(xì)胞誘導(dǎo)，分別于誘導(dǎo)的第1W、2W、3W進(jìn)行形態(tài)學(xué)觀察、甲苯胺藍(lán)染色、阿利新藍(lán)染色和Ⅱ型膠原（ColⅡ）的免疫組化染色。 【結(jié)果】所獲取細(xì)胞呈類成纖維細(xì)胞樣貼壁快速生長(zhǎng)，生長(zhǎng)曲線呈比較典型的“S”型；傳代后生長(zhǎng)、增殖穩(wěn)定可達(dá)20代以上；細(xì)胞表面抗原CD34、CD45呈陰性反應(yīng)，CD44、CD29呈陽(yáng)性反應(yīng)，陽(yáng)性率達(dá)97%以上；在TGF-β3的誘導(dǎo)下，細(xì)胞形態(tài)逐漸變圓，甲苯胺藍(lán)染色、阿利新藍(lán)染色和Ⅱ型膠原的免疫組化染色均呈強(qiáng)陽(yáng)性，而對(duì)照組和空白組則不明顯。 【結(jié)論】初步鑒定成功分離得到了ADSCs，細(xì)胞體外增殖穩(wěn)定快速，并且具有較好的成軟骨分化的潛能，含10ng/mlTGF-β3濃度的誘導(dǎo)液更有利于ADSCs向軟骨細(xì)胞分化。ADSCs有望成為理想的種子細(xì)胞應(yīng)用于軟骨組織工程以修復(fù)軟骨損傷。
[Abstract]:[objective] to investigate the isolation, culture and identification of rabbit adipose derived stem cells (rabbitadipose-derivedstemcells,RADSCs), and to explore the feasibility of promoting the differentiation of RADSCs into chondrocytes with different concentrations of transforming growth factor 尾 3 (TransformingGrowthFactor- 尾 3 TGF- 尾 3). To provide a new idea for the selection of seed cells in cartilage tissue engineering. [methods] Adipose tissue of inguinal region or neck and back of adult white rabbits were obtained by enzyme digestion and density gradient centrifugation, primary and passage culture were carried out, cell changes were observed and cell growth curve was drawn. Flow cytometry and immunofluorescence techniques were used to identify the CD molecules on the cell surface. The third generation cells were induced by chondroblasts in vitro. Morphological observation, toluidine blue staining, alimine blue staining and type 鈪

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