Study on the culture of rat myocardium microvascular endothelial cells and microarray analysis

AIM: To study the cytological characteristics of rat myocardium microvascular endothelial cells (RMMVEC) by microarray. METHODS: The RMMVEC were cultured by the method of planting myocardium tissue. The morphology of RMMVEC was studied by light and electronic microscopy. Its molecular markers were observed by immunocytochemistry. Cell proliferation kinetic was analyzed by counting the number of cells. The gene expression of the RMMVEC was studied by endothelial cell biology gene microarray and compared the change of gene expression among the cultured cells of primary, 2nd and 5th passage. RESULTS: The RMMVEC showed morphological characteristics of microvascular endothelial cells (MVEC): growing in a cobblestone pattern, forming tube-like structure or capillary network and having microvilli on cell surface. At the same time, the RMMVEC showed positive staining for vWF, CD34, CD31, CD105 and Tie-2. Gene microarray analysis indicated expression of VEGFR, ICAM-1, VCAM-1, angiopoietin1, PECAM1 (CD31) and other genes closely related to microvascular endothelial functions at relatively high level. But in cultured cells of 5th passage the characteristic gene expression of microvascular endothelial cells disappeared. CONCLUSION: The RMMVEC cultured by this method possess typical characteristics of MVEC. The cytological characteristics are steady in the cultured cells of primary and 2nd passage. It can be utilized to study the mechanisms of some cardiovascular diseases.