【作者】 宋晓平； 窦忠英； 雷安民； 华进联； 杨学义； 马勇江；

【Author】 SONG Xiao-ping, DOU -yin Zhong g, LEI An-min, HUA Jin-Han, YANG Xue-yi,MA Yong-jiang(Shaanxi Centre of Stem Cells Engineering & Technology, Northwest Sci-Tech University of Agriculture and Forestry,Yangling, Shaanxi 712100, China)

【机构】 西北农林科技大学陕西省干细胞工程技术研究中心；

【摘要】 新生1～3d的小鼠50只拉颈处死,无菌采取颅盖骨。在室温条件下,用0.25%胰蛋白酶和0.04%EDTA组成的细胞消化液,磁力搅拌消化30min,充分除去骨片周围及骨缝处的结缔组织,再将小骨片按常规植块法培养8～10d,差速黏附贴壁纯化成骨细胞,并在传代培养2d后收集培养液,制备用于胚胎干细胞诱导分化成骨细胞的条件培养基。更多还原

【Abstract】 Fifty neonatal mice, 1-3 day-old, were killed by cervical dislocation, and collected their calvaria under sterile condition. To sufficiently remove connective tissue surrounded the calvaria and at the cranial sutures, the calvaria were treated with 0.25% trypsin and 0.04% EDTA in saline under being mixed round by magnetic force at room temperature for 30 min. And then small pieces of calvaria were cultured by common expant culture for 8-10d. Secondary culture was carried out with different attaching pace between osteoblasts and fibroblasts for purifying osteoblasts, and two days later this medium was collected to prepare the conditioned medium for being applied to induce differentiation of embryonic stem cells.更多还原