| When stroke happens,the damage of cerebral vessels is the main pathological change.The vascular anatomic structure are unble to completly recovery by treatment at present to supply the blood of ischemia half dark belt,so forming blood vessels in ischemic area becomes even more significant.There are two kinds of situation to form vascular called agionenesis and vasculogenesis. Agionenesis gives rise to new capillaries budding from the original vesseles,while vasculogenesis forms new blood vessels at avascular zone,just as the EPCs come together to form new blood vessels at damaged or ischmic tissue selectively. When the body suffer from ischemia and hypoxia,these two style complement with each other to promote the formation of new blood vessels.The research on EPCs in the field of ischemic diseases has become more and more. Our study was designed to isolate EPCs from rat bone marrow and peripheral blood and identify them,and found the characteristics and differences between them.Mononuclear cells were obtained by using density gradient centrifugation and adherence screening method from bone marrow and peripheral blood of rat and induct them.Finally observed and recorded the biological characteristics of adherent cells. EPCs specific surface markers CD133, CD34, KDR is selected and detected by immunofluorescence. Flow cytometry was used to detect KDR, CD34, Phagocytosis experiments was carryed to further identify cells.Experimental results;(1) EPCs can be isolated from bone marrow and peripheral blood of rat.(2) The growth of EPCs from bone marrow and peripheral blood in common:1. Delayed attachment.2. A variety of cell morphology:round cells, spindle cells, "Blood Island" colony;gathered round the central cell with the cluster radial cells surrounding,cord-like, tubular, cobblestone. (3)The difference of biological characteristics of EPCs from Bone Marrow and PB:1.The number of cell colony was significantly lower in PB than in BM; 2.BMC attached more than in PB,but had different forms.3.EPCs from BM had great activity,and can store and proliferate many generations.They grew and connected together, formed vascular cavity,while EPCs in PB had lower activity,and lasted about 1 month to become circular and scarce, then apotosis and disappeared.(4)Attached cells were positive for CD34, KDR and CD133 by immunofluo-rescence,(5) Cells were positive for markers CD34, KDR, the level are 69.0%,84.8%(BM),5.4%。32.7%(PB) respectively;(6) Cells can swallow ac-LDL, bound with UEA-1, showed that the cells we obtained had complete function of EPCsEPCs are kinds of precursor cells that can differentiate into a vascular endothelia. It not only involved in embryonic blood vessel formation, but also played founds in adult individuals with angiogenesis and angiogenesis closely related stem cells, They played an important role in repairing vascular and collateral vascular network,had founctions in promoting angiogenesis in ischemic vascular regeneration, and furthering nerve regeneration, predicting the development of cerebral ischemia and so on.Transplantation of EPCs are used in the treatment of vascular disease recently. of EPCs used as seed cells to angiogenesis is more significant, along with gene therapy can be better. For the purpose of clinical applications, we isolated and culcured EPCs from bone marrow and peripheral blood of rat based on animal experiments, and searched a stable, practical method to isolate, cultivate and identificate EPCs in vitro.Our aim is to explore the best EPCs amplification system in order to provide sufficient better quantities source of EPCs that can be used in cinical.
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