接种密度对人脐静脉间充质干细胞原代培养的影响

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

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5×10^5×10^6/cm^2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。

Objective： To explore the influence of different implantation density on the primary culture of mesenehymal 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-indueed culture were performed on those cells. The expressions of bone tubercula were examined. Results： It was reasonable that hUV- MSCs were plated at 5 × 10^5- 1 × 10^6/cm^2 cell density. After osteogenesis induced by some chemicals （dexamethasone,β-glycerphosphoric sodium and vitamin C） the bone tubereula were formed, Condnsion：Under optimal conditions, lots of autograft seeding cells can be obtained in short time and tissue engineering bone can be eontrueted in vitro.