Optimization of Animal Sera-Free Culture Condition for Generation and Expansion of Human Cardiosphere-Derived Cells

Background: Preclinical studies have introduced cardiac stem/progenitor cells (CSCs) as a promising cell candidate for cell therapy of heart diseases. CSCs can be isolated from myocardial biopsies using various protocols, expanded in vitro and transplanted back to the patients. One of the most important issues regarding the clinical usage of cells is the choice of suitable humanized culture supplements to replace commonly used animal-derived products including fetal bovine serum (FBS) or fetal calf serum (FCS).Methods and Materials: In order to find the optimal FBS substitute, human myocardial samples were cultured as explants in media supplemented with one of these different blood products: FBS, human serum (HS), human plasma (HP) or platelet lysate (PL). The out-grown cells were cultured in suspension to generate cardiospheres and then plated to expand as cardiosphere-derived cells (CDCs). The effect of culture media on the process of CDC generation and culture was evaluated in terms of morphology and cell growth.Results: Among the examined humanized agents, CDCs were only generated and expanded in medium supplemented with HS. Furthermore, they had normal karyotype and expressed CSC associated surface markers but not endothelial and hematopoietic markers. Moreover, cultured CDCs in HS inhibited the proliferation of induced lymphocytes in vitro which might highlight the immuno-modulatory feature of these cells.Conclusion: Taken together, our data exhibited the superiority of HS compared to other tested human blood products for CDC culture which can be suggested for cell culture set up of cardiac clinical studies.Background: Preclinical studies have introduced cardiac stem/progenitor cells (CSCs) as a promising cell candidate for cell therapy of heart diseases. CSCs can be isolated from myocardial biopsies using various protocols, expanded in vitro and transplanted back to the patients. One of the most important issues regarding the clinical usage of cells is the choice of suitable humanized culture supplements to replace commonly used animal-derived products including fetal bovine serum (FBS) or fetal calf serum (FCS). Methods and Materials: In order to find the optimal FBS substitute, human myocardial samples were cultured as explants in media supplemented with one of these different blood products: FBS, human serum (HS), human plasma (HP) or platelet lysate (PL). The out-grown cells were cultured in suspension to generate cardiospheres and then plated to expand as cardiosphere-derived cells (CDCs). The effect of culture media on the process of CDC generation and culture was evaluated in terms of morphology and cell growth. Results: Among the examined humanized agents, CDCs were only generated and expanded in medium supplemented with HS. Furthermore, they had normal karyotype and expressed CSC associated surface markers but not endothelial and hematopoietic markers. Moreover, cultured CDCs in HS inhibited the proliferation of induced lymphocytes in vitro which might highlight the immuno-modulatory feature of these cells. Conclusion: Taken together, our data exhibited the superiority of HS compared to other tested human blood products for CDC culture which can be suggested for cell culture set up of cardiac clinical studies.

کلیدواژه ها:

Progenitor Cells ، Culture Medium ، Human Serum ، Cell Therapy