Application Of Porcine Platelet Lysate In Buffalo Mesenchymal Stem Cell Culture In Vitro

Fetal bovine serum(Fetal bovine serum,FBS),as a routine cell culture supplement,plays an important role in cell acquisition and in vitro culture applications.However,the price of FBS is relatively high and the cell culture effects of different brands of FBS are different.At the same time,Platelet lysate(Platelet lysate,PL)can also be used as a cell culture additive.If pigs are used as the source animal for platelet lysate,the production cost relatively low.In order to explore whether Porcine platelet lysate(Porcine platelet lysate,PPL)can replace FBS as a cell culture additive,PPL and FBS were used to culture Jianghuai buffalo bone marrow mesenchymal stem cells,and the growth performance and induced differentiation of buffalo cells in two culture conditions were compared.Objective: To culture Jianghuai buffalo bone marrow mesenchymal stem cells(BMSCs)with fetal bovine serum and porcine platelet lysate,and to compare the growth of buffalo bone marrow mesenchymal stem cells in PPL and FBS.Cell characteristics in culture supplements,providing a theoretical basis for platelet lysate to replace serum for culturing cells.Methods: Fresh anticoagulated pig blood was collected to prepare pig platelet lysate.Jianghuai buffalo bone marrow mesenchymal stem cells were cultured in medium containing 10% PPL and FBS,and the cell status was observed under a microscope.The proliferation efficiency of the third passage cells under the two culture conditions for 7 days was measured by the cck-8 cell detection kit,and the cell growth curve was drawn.The expression of surface-specific antigens of buffalo BMSCs under two culture conditions was identified by immunofluorescence staining.Finally,the adipogenic,osteogenic,and chondrogenic differentiation properties of buffalo BMSCs cultured under PPL and FBS conditions were determined.Results:(1)Porcine platelet lysate was successfully prepared;(2)The cells cultured with 10% PPL and FBS conformed to the morphological characteristics of mesenchymal stem cells,and there was no significant difference in the morphology of cells cultured in PPL and FBS;(3)The third passage cell growth curve results showed that the proliferation rate of cells cultured with FBS was slightly higher than that of PPL;(4)The results of cell immunofluorescence showed that the staining results of buffalo BMSCs under the two culture conditions were relatively consistent;(5)Cells In vitro induction test showed that buffalo BMSCs maintained the ability to induce differentiation under PPL and FBS culture conditions,and the differentiation performance of cells was slightly different under different culture conditions.Conclusion:(1)PPL was successfully prepared,and the cells were cultured in PPL to maintain biological characteristics;(2)Compared with FBS,PPL did not significantly change the biological characteristics of cells after being used as a cell culture additive.The cells were all pluripotent,which provided a basic data reference for PL to replace FBS to culture cells;(3)PPL and FBS showed their respective culture advantages in the process of inducing differentiation.This experiment compared the cell biological characteristics of Jianghuai buffalo BMSCs within two culture conditions and provided basic data support for the future use of PPL instead of FBS to culture cells.