接种密度对人脐静脉间充质干细胞原代培养的影响

目的:探讨不同接种密度对人脐静脉间充质干细胞(hUV-MSCs)原代培养的影响,从而确定原代培养hUV-MSCs的适宜条件。方法:分离人脐静脉内皮及内皮下层细胞,按不同接种密度分组进行原代培养,记录原代培养时间,然后进行传代和成骨诱导培养,并检测细胞钙结节的表达情况。结果:hUV-MSCs原代分离培养消化30min,种植细胞密度为5×105~1×106/cm2的方法较合理。经化学药物(如地塞米松、β-甘油磷酸、维生素C)成骨诱导后,可形成钙结节。结论:合理hUV-MSCs原代培养方法,可以快速获得最大量的自体种子细胞,从而为组织工程化骨的体外构建创造条件。

Influence of Implantation Density on Primary Culture of Mesenchymal Stem Cells From Human Umbilical Vein

Objective:To explore the influence of different implantation density on the primary culture of mesenchymal stem cells from human umbilical vein(hUV-MSCs) and ensure the optimum conditions for their primary culture.Methods:The isolated human umbilical vein cells were divided into groups according to different implantation density and then were primarily cultured.The culture time was recorded.Then,secondry culture and osteogenesis-induced culture were performed on those cells.The expressions of bone tubercula were examined.Results: It was reasonable that hUV-MSCs were plated at 5×105～1×106/cm2 cell density.After osteogenesis induced by some chemicals(dexamethasone,β-glycerphosphoric sodium and vitamin C) the bone tubercula were formed.Conclusion:Under optimal conditions,lots of autograft seeding cells can be obtained in short time and tissue engineering bone can be contructed in vitro.