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2.9. Cell culture Human mesenchymal stem cells (hMSCs) were a kind gift donated by the Second Affiliated Hospital of Sun Yat-Sen University and were propagated in Dulbecco’s Modified Eagle’s Medium (DMEM) with supplements of 1.5 mg$ml-1 sodium bicarbonate,4.5 mg ml-1 glucose, 10% (v/v) fetal bovine serum (FBS), 3 mg ml-1 4-(2- hydroxyerhyl)piperazine-1-erhanesulfonic acid (HEPES). The cells were kept in a humidified incubator at 37℃and5%CO2, and themediumwas changed every 3 days. 2.10. Cell seeding on the scaffolds PLGA/PHBV scaffolds were cut into disks of 6mmin diameter and 2mmin height and sterilized by exposing to gamma radiation (15 kGy). The pure PLGA scaffolds of the same size as that of PLGA/PHBV were immersed in 75% (v/v) ethanol aqueous solution for 2 h and followed by ultraviolet radiation for 30 min to sterilize. All the scaffolds were prewet in the DMEM solution for 24 h. Fifteen microlitres of passage 6 cells in suspension (7.5×104 cells/well) were seeded on each scaffold. 2 h later, 700 μl of culture medium was added into each well. The cells/scaffold constructs were incubated at 37℃ in a humidified incubator of 5% CO2 for pre-set days. |
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转身丶忘不掉: 金币+20, 翻译EPI+1, ★★★★★最佳答案 2014-12-07 13:41:10
转身丶忘不掉: 金币+20, 翻译EPI+1, ★★★★★最佳答案 2014-12-07 13:41:10
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细胞培养 人类间充质干细胞(hMSCs)是一种礼物由中山大学第二附属医院的捐赠和在Dulbecco修改鹰的传播媒介(DMEM)补充1.5毫克ml-1美元碳酸氢钠,4.5毫克ml-1葡萄糖,10%(v / v)胎牛血清的边后卫,3毫克ml-1 4 -(2 - hydroxyerhyl)piperazine-1-erhanesulfonic酸(玫瑰)。这些细胞被保存在一个湿润孵化器在37℃and5%CO2,和themediumwas每3天换一次。 2.10。细胞支架上播种 PLGA / PHBV支架被切成磁盘6 mmin直径和2 mmin高度和消毒暴露伽马辐射(15 kGy的)。相同大小的纯PLGA支架的PLGA / PHBV沉浸在75%(v / v)乙醇水溶液为2 h,紧随其后的是紫外线消毒30分钟。所有的支架都是两高一低的DMEM 24 h。15毫升通道解决方案6细胞悬浮(7.5×104细胞/)被播种在每个支架。2 h后,700μl培养基添加到每个。细胞/支架结构在37℃湿润孵化器孵化预设天5%的二氧化碳。 |
2楼2014-12-05 15:47:21