不同浓度胰岛素样生长因子-1和转化生长因子β—3对人肌腱细胞存活和胶原合成的影响  

The effects of insulin-like growth factor 1 and transforming growth factor β-3 at various concentration on tenocyte survival and collagen formation

作　　者：邱轶伟[1] 朱理玮[1] 张鑫[1] 张鹏[1] 

机构地区：[1]天津医科大学总医院普通外科,300052

出　　处：《中华外科杂志》2012年第8期744-747,共4页Chinese Journal of Surgery

基　　金：英国牛津大学Nuffield骨科中心,骨科、风湿科以及肌肉关节科学系NIHR-BRU基金资助项目;天滓市卫生局科研基金资助项目（2011KZ118）;天津市教委科研基金资助项目（20110121）

摘　　要：目的为肌腱组织工程筛选出一种优化的人肌腱细胞培养基，即使在不添加胎牛血清（fetalbovineserum，FBS）的条件下，通过优化培养基构成维持其存活并分化。方法在α—MEM培养基中，加入最佳组合的生长因子（growthfactors，GF），同时加入不同浓度的FBS（0、1％、5％和10％）和不同浓度的IGF-1（0、10、50ng／ml），TGFβ—3（0、1、10ng／ml），用拉马拉蓝染色法检测人肌腱细胞的生存情况、增殖速度，用天狼星红定量胶原合成。结果在无FBS情况下，含有10ng／mlTGFβ—3和50ng／mlIGF-1两种生长因子的α-MEM培养基中培养14d后，人肌腱细胞依旧存活（6228．68±43．87），高于其他实验组，但低于10％FBS对照组（13576．74±286．75，t=41．29，P〈0．05），胶原合成量[（0．322±0．003）ng]明显大于未添加生长因子组（t=4．13～5．93，P〈0．05）。结论添加50ng／mlIGF-1和10ng／mlTGFβ—3优化的α-MEM培养基可维持人肌腱细胞存活长达14d并促进肌腱细胞分化，而不必添加FBS。Objective To optimize the culture media by adding the growth factors required to maintain tenocytes survival and promote their differentiation without fetal bovine serum （FBS） supplementation, in order for the approach to be used for any future tendon tissue engineering. Methods The human tenoeytes were cultured in α-MEM media by adding FBS at various concentrations and supplementing both insulin-like growth factor 1 （IGF-1） and transforming growth factor β-3 （TGFβ-3）. A number of growth factors were selected that could support tenocytes expansion at reduced differentiated state with the minimum FBS. By employing fractional factorial design, different treatment groups went through AlamarBlueTM tests to evaluate the cell number growth whilst collagen quantification by real time RT-PCR technique and tenocyte differentiation were also studied. Results The tenocytes cultured for 14 days with 0% FBS, 50 ng/ml IGF-1 and 10 ng/ml TGFβ-3 maintained survival over 14 days, the Cell count were 6228.68±43.87. They were higher than the other experimental groups, but less than 10% FBS control group （ 13 576. 74 ± 286. 75, t = 41.29, P 〈 0. 05 ）. The tenocytes cultured in the treated group also showed enhanced collagen synthesis （ （0. 322 ± 0. 003 ） ng, t = 4. 13-5.93, P 〈 0. 05 ） . Conclusion These findings have shown for the first time that human tenoeytes could be maintained survival for a long period of time in the culture media without FBS, having this approach a suitable one for tendon tissue engineering.

关 键 词：组织工程 细胞存活 胶原 胰岛素样生长因子I 转化生长因子Β3 

分 类 号：R686[医药卫生—骨科学]