【摘要】：研究目的 探討腱組織提取物對(duì)ADSC增殖和向肌腱細(xì)胞方向分化的影響,從而為肌腱組織工程及其最終應(yīng)用于臨床提供理論依據(jù)。 研究方法 從3周齡SD大鼠腹股溝脂肪墊分離出脂肪組織,通過(guò)Ⅰ型膠原酶消化法分離干細(xì)胞,接種于含有胎牛血清的DMEM培養(yǎng)基中分離傳代。每日以倒置顯微鏡觀察細(xì)胞形態(tài)及生長(zhǎng)情況,MTT法繪制ADSC生長(zhǎng)曲線。經(jīng)鑒定后,取第三代ADSC分成實(shí)驗(yàn)組和對(duì)照組2組,利用腱組織提取物處理實(shí)驗(yàn)組ADSC, Hank's液處理對(duì)照組細(xì)胞后繼續(xù)培養(yǎng)觀察。采用MTT比色法進(jìn)行增殖活性比較,Ⅰ型膠原免疫細(xì)胞化學(xué)染色檢測(cè)成肌腱分化情況以及RT-PCR檢測(cè)肌腱發(fā)育相關(guān)基因(CollagenⅠ、CollagenⅢ基因)的表達(dá)。 研究結(jié)果 原代細(xì)胞接種4-6小時(shí)后即可見(jiàn)沉降貼壁,7-8天左右可見(jiàn)大量增殖,按一定方向性排列,呈漩渦狀或束狀,傳代后細(xì)胞增殖迅速,生長(zhǎng)穩(wěn)定。腱組織提取物誘導(dǎo)后,MTT實(shí)驗(yàn)顯示ADSC增殖活性并未發(fā)生明顯改變,與對(duì)照組間沒(méi)有統(tǒng)計(jì)學(xué)差異(p0.05)。行Ⅰ型膠原免疫細(xì)胞化學(xué)染色顯示：實(shí)驗(yàn)組為陽(yáng)性,對(duì)照組陰性；RT-PCR檢測(cè)顯示肌腱發(fā)育相關(guān)基因(CollagenⅠ、CollagenⅢ基因)表達(dá)明顯增加(p0.01)。 研究結(jié)論 1.腱組織提取物不能改變大鼠ADSC的增殖活性 2.腱組織提取物可以促進(jìn)大鼠ADSC向肌腱細(xì)胞方向分化
[Abstract]:Objective to investigate the effect of tendon tissue extract on the proliferation and differentiation of ADSC in the direction of tendon cells, so as to provide theoretical basis for tendon tissue engineering and its application in clinical practice. Methods Adipose tissue was isolated from the inguinal fat pad of 3-week-old SD rats. Stem cells were isolated by type I collagenase digestion method. The stem cells were isolated and subcultured in DMEM medium containing fetal bovine serum. The morphology and growth of cells were observed by inverted microscope daily, and ADSC growth curve was drawn by MTT method. After identification, the third generation of ADSC was divided into two groups: the experimental group and the control group. The cells of the control group were treated with the extract of tendon tissue and treated with the solution of ADSC, Hank's. The proliferative activity was compared by MTT colorimetry, the differentiation of tendon was detected by type I collagen immunocytochemical staining, and the expression of Collagen 鈪，

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