Research On Isolation, Culture And Differentiation Potential Of Pulmomary Mesenchymal Stem Cells In Beijing Fatty Chicken

We study the isolation, culture and differentiation potential of Pulmomary mesenehymal stem cells in Beijing Fatty Chicken.This research aimed to establish an optimal in vitro isolation and amplification system for Beijing Fatty Chicken pulmomary mesenehymal stem cells (PMSCs), and to thereby laying a technical foundation for future pertinent researches.Lung tissue were isolated from 9-14-day-old chicken embryos, disassociated with collagenaseⅣ, and the cells were purified with percoll. Identification was carried out by immunohistochemisty and RT-PCR. Proliferative capacities in three different culture systems were compared as well. We also explored the differentiation potency of PMSCs. The results were following:1. Proliferative capacities in three different culture systems were compared as well. Lung tissue were isolated from 9-14-day-old chicken embryos.The best method is that Lung tissue were isolated from 9-14-day-old chicken embryos, disassociated with collagenaseⅣ, and the cells were purified with percoll.The suitable culture condition for PMSCs was DMEM/F-12+10%FBS+2mM glutamine+100 IU/ml penicillin+100μg/ml streptomycin10ng/ml bFGF.2. The different passages growth curves of PMSCs showed typical"S"shape. The cell growth experienced the processes of latentperiod, logarithmic phase and plateau phase. The cells were sub-cultured to the 25th passage. The cells sub-cultured to the 25th passage showed representative senescent appearance, and could not be sub-culture anymore.3. We studied the influences of freezing conservation and recovery on cell viability. The results showed that cryopreservation did affect cell viability in P3,P15,P21cells, and the cell viability decline .This demonstrated that the anthropic factor should be considered and the composition of frozen stock solution should be optimized.4. We identified the PMSCs by the means of immunohistochemisty and RT-PCR .The results showed that PMSCs expressed the MSCs markers of CD29, CD44,CD71,CD73and Vimentin while did not express endothelial cells specific markerCD34. It showed that they were indeed Beijing Fatty chicken PMSCs.5. Beijing Fatty Chicken PMSCs could be induced to differentiate into osteoblasts, adipocytes, cardiac myocytes in the condition of appropriate effect,which identified by the means of immunohistochemisty and RT-PCR.This provde that Beijing fatty chicken PMSCs have the differentiation potency.