Complex Fang Shuangdan Active Component Screening And Bone Marrow Mesenchymal Stem Cell Culture Studies,

. Heart and vascular desease is one of the main cause of the death of human.Traditional Chinese medicine Shuangdan Formula(Salvia miltiorrhiza Bge andPaenoina Suffruticosa Andr) has excellent effect on incurring the heart andvascular desease. Bone marrow mesenchymal stem cells(BMMSCs) are pluripotentstem cells, they can be induced to differentiate to myocardial cell, moreover theyhave no ethic problem, so transplant BMMSCs provide a wide prospect to cure heartdesease. Because Traditional Chinese Medicine is a complex chemical substancesystem, it is very difficult to illustrate the effect compounds. For the reason of fewbone marrows in the young rats, the isolation and purification of the rat BMMSCsare very difficult.In this paper, under the guide of the effect direct leading compound screening,Shuangdan Formula 51 fractions were screened by detected the Lactic aciddehydronase activity of myocardial cell imitated myocardial infarction andreperfusion. 3 fractions of the Salvia miltiorrhiza Bge were screened effective, 2of them were ester dissolving fractions and the other one was H2O dissolvingfraction. 6 fractions of the Paenoina Suffruticosa Andr were screenedeffective, 4 of them were ester dissolving fractions and the other 2 wereH2O dissolving fraction. The main compounds the 9 fractions were Salviamiltiorrhiza aldehyde, Salvia miltiorrhiza ketone, Salvia miltiorrhizaalcohol and others.In this paper, Rat BMMSCs were isolated, purified and identified.BMMSCs were isolated by Percoll density gradient centrifugation andpurified by different adherence speed. Then BMMSCs were identified andcharacterized though seven different CD surface antigens by flow cytometryand laser confocal technology. CD29, CD44, CD105, CD166 expressedpositively , while CD14, CD34, CD45 just in contrast. Three mark genes ofstem cells were identified by RT-PCR, nestin and NST were high positiveexpression and Oct-4 was low positive expression. The results accorded tothe expression of genes and antigens on the surface of BMMSCs, this couldconclude that the cells extracted from rat marrow were BMMSCs. This easyand useful protocol to purify the BMMSCs could supply seminal cells for cell andtissue engineering, and it also has great significance in basic research and clinicalapplication.