低氧时脂肪间充质干细胞如何向跟腱细胞分化

背景：脂肪间充质干细胞在跟腱组织工程再生领域越来越受到重视，研究其诱导分化的有利环境(氧体积分数)尤为必要。 目的：将脂肪间充质干细胞与跟腱细胞在不同氧体积分数条件下间接共培养，比较氧体积分数对脂肪间充质干细胞向跟腱细胞分化能力的影响。 方法：无菌条件下获取SD大鼠跟腱细胞。SD大鼠脂肪间充质干细胞直接购买，经培养传至第1代后，与跟腱细胞一起在常氧(体积分数20%)和低氧(体积分数2%)条件下经Transwell间接共培养体系共培养。在培养7，14和21 d，实时荧光定量PCR检测跟腱细胞的特异指标胶原蛋白Ⅰ、胶原蛋白Ⅲ，Tenomodulin，Thrombospondin-4，Scleraxis基因相对表达量，免疫荧光染色法检测胶原蛋白Ⅰ和Thrombospondin-4蛋白表达量。 结果与结论：脂肪间充质干细胞与跟腱细胞经间接共培养后，低氧组检测到的跟腱细胞的相关特异指标在基因和蛋白水平上表达水平均高于常氧组。提示氧体积分数可显著影响脂肪间充质干细胞向跟腱细胞分化的潜能，低氧是脂肪间充质干细胞向跟腱细胞分化的有利条件之一。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：

Tenogenic differentiation of adipose-derived stem cells in hypoxia

BACKGROUND:Adipose-derived mesenchymal stem cells are gaining widespread interest in the Achilles tendon tissue engineering and regeneration, and an enabling environment (oxygen concentration) for cell induction and differentiation is particularly necessary. OBJECTIVE:To co-culture adipose-derived mesenchymal stem cells with primary tenocytes in standard culture condition (20% O2 tension) and in an atmosphere of reduced oxygen (2% O2 tension) in order to determine whether the two conditions differ in their effect on tenogenic differentiation. METHODS:Tenocytes were isolated via serial expansion in culture from several Sprague-Dawley rats’ Achilles tendons. Adipose-derived mesenchymal stem cells were purchased. After one passage, adipose-derived mesenchymal stem cells were indirectly co-cultured with tenocytes in standard culture condition (20% O2 tension) and in an atmosphere of reduced oxygen (2% O2 tension). Collagen 1, collagen 3, Tenomodulin, Thrombospondin-4, Scleraxis levels were compared for each culture condition at 7, 14 and 21 days following co-culturing. Immunofluorescence staining was performed to evaluate production of collagen 1 and Thrombospondin-4. RESULTS AND CONCLUSION:Following indirect co-culturing, hypoxic differentiated adipose-derived mesenchymal stem cells expressed higher levels of tendon-related genes and proteins than normoxic controls, which suggest that oxygen levels can significantly affect tenogenic differentiation, and hypoxia is advantageous for efficient differentiation of adipose-derived mesenchymal stem cells in vitro for tendon tissue engineering.