In vitro Expansion and Differentiation of Hematopoietic Stem Cells into Platelets.

The aim: This study aimed to collect mobilized HSCs by aphaeresis and culturing the cells under effect of various growth factors cocktails to generate mature megakaryocytes and platelets. Material and Methods: The present study was conducted on 10 subjects divided into: (group 1) 5 patients of liver disease with thrombocytopenia and (group 2) 5 healthy control. The culture protocol lasted for 14 days, the process of megakaryopoiesis was characterized by flow cytometry using fluorochrome-conjugated antibodies against CD61, CD34 and CD42. Mature MKs were observed as larger cells in culture using a phase contrast inverted microscope. Results: HSCs cultured for 14 days in growth factors cocktail revealed weak expression of CD34 and positive expression of both CD61 and CD42 for both CLD and control groups compared to those before culturing. Moreover, inverted microscope study on the 14th day revealed the presence of mature MK which can be observed as larger cells in the culture. Conclusion: The present study revealed successful generation of MK and platelets having CD61+ve and CD42+ve surface markers from differentiation of HSCs derived from peripheral blood which is an easy accessible source with no ethical problems. This study also demonstrated that CLD doesn't affect HSCs differentiation into MK and platelets. These results together with those of previous studies constitute important implications for the development of regenerative medicine in patients suffering from thrombocytopenia.