Research of endothelial cells induced from rabbit marrow mononuelear cells in vitro / 中国矫形外科杂志

ABSTRACT

[Objective]To research the efficient way of isolating and proliferating of induced endothelial cells(ECs) ,measure the number of the ECs and observe the main biological characteristic of ECs m vitro during the passage cultivation.[Method]The marrow mononuclear cells(MNCs) in rabbit marrow were isolated by the gradiem centrifugation and cultivated to isolate and induce ECs. The efficiency of isolating and harvesting ECs was measured by the number of the first passage ECs from every 10~6 of the MNCs. The changes of ECs morphology, growth and proliferation were observed during the passage cultivation. The immunohistochemical staining of CD31 and vWF and function of ECs were checked.[Result]After cultivated at 1?10~6/cm~2 MNCs for 7～8 d, the first passage ECs were harvested. The number of the first and the second passage ECs had a remarkable statistical positive correlation with the number of the MNCs from one rabbit. After cultivated for 7～8 d, the first passage ECs of 7.086?0.75?10~4 in number and the second passage ECs of 53.20?6.47?10~4 in number were harvested from every 10~6 MNCs on average;ECs, which were monolayer arrayed to form cobblelike shape,The cells of 2nd passage showed positive staining of CD31 and vWF, and positive labeling of DiI-LDL and UEA.[Conclusion]The period from cultivating to harvesting ECs is shortened at the density of 1?10~6/cm~2 MNCs. It is feasible that the efficiency of isolating and harvesting ECs could be measured by the number of the first or the second passage ECs from every 10~6 MNCs. The induced ECs can act as seeding cells in bone tissue engineering in future.