【摘要】：目的對(duì)從膝關(guān)節(jié)軟骨損傷區(qū)分離出的細(xì)胞進(jìn)行鑒定,并對(duì)其生長(zhǎng)活性進(jìn)行研究,以解決自體軟骨細(xì)胞移植中軟骨細(xì)胞來(lái)源不足的問(wèn)題。方法通過(guò)膝關(guān)節(jié)鏡,在軟骨損傷區(qū)取少量損傷軟骨組織,不含正常軟骨及軟骨下骨為實(shí)驗(yàn)組;取正常軟骨組織為對(duì)照組。在胰蛋白酶和Ⅱ型膠原酶作用下從實(shí)驗(yàn)組中提取細(xì)胞,對(duì)照組中提取正常軟骨細(xì)胞。實(shí)驗(yàn)組細(xì)胞培養(yǎng)貼壁后在倒置相差顯微鏡下觀察細(xì)胞的形態(tài)是否與對(duì)照組軟骨細(xì)胞一致。實(shí)驗(yàn)組細(xì)胞行甲苯胺藍(lán)染色鑒定,與對(duì)照組對(duì)比;噻唑藍(lán)(MTT)法比較兩組細(xì)胞生長(zhǎng)活性;以具有專(zhuān)利知識(shí)產(chǎn)權(quán)的Ⅰ/Ⅱ型復(fù)合膠原膜為生長(zhǎng)載體,培養(yǎng)實(shí)驗(yàn)組和對(duì)照組第3代軟骨細(xì)胞,在電鏡下觀察兩組生長(zhǎng)活性。結(jié)果從少量損傷的軟骨組織中提取的細(xì)胞為活性正常的軟骨細(xì)胞,其特征及生長(zhǎng)活性與對(duì)照組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論軟骨損傷過(guò)程中軟骨細(xì)胞未發(fā)生變性,少量損傷的軟骨組織仍可獲得適量軟骨細(xì)胞,MTT法和電鏡掃描表明,其生長(zhǎng)活性符合自體軟骨細(xì)胞移植的要求,無(wú)需取正常軟骨組織作為細(xì)胞來(lái)源。
[Abstract]:Objective to identify the cells isolated from cartilage injury area of knee joint and to study their growth activity in order to solve the shortage of chondrocytes in autologous chondrocyte transplantation. Methods through knee arthroscopy, a small amount of injured cartilage tissue was taken from the injured area of cartilage, without normal cartilage and subchondral bone as experimental group, and normal cartilage tissue was taken as control group. The normal chondrocytes were extracted from the experimental group and the control group under the action of trypsin and collagenase. The morphology of the cells in the experimental group was observed under inverted phase contrast microscope after adherent to the wall and was consistent with the chondrocytes of the control group. The cells of the experimental group were identified by toluidine blue staining and compared with the control group, and the growth activity of the two groups was compared by (MTT) assay. The third passage chondrocytes of the experimental group and the control group were cultured with the type 鈪，

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