Blood Res  
Human platelet lysate efficiency, stability, and optimal heparin concentration required in culture of mammalian cells
Hoda E. Mohamed1, Mervat E. Asker1, Nahla Shehata2, Akram M. El Habab2
1Department of Biochemistry, Faculty of Pharmacy, Zagazig University, Egypt; 2National Organization for Research and Control of Biologicals, Egypt
Correspondence to: Akram M. El Habab
National Organization for Research and Control of Biologicals, Giza, Egypt
E-mail: akram.elhabab@yahoo.com
Published online: March 13, 2020.
© The Korean Journal of Hematology. All rights reserved.

Abstract
Background: Fetal bovine serum (FBS) has been used to support the growth and proliferation of mammalian cells for decades.
Methods: In this study, human platelet lysate (HPL) derived from activated human platelets was evaluated as an alternative to FBS due to the associated risk factors. To evaluate the efficiency of the preparation process, platelet count was performed before and after activation. The concentrations of several growth factors and proteins were measured to investigate HPL efficiency. HPL stability was studied at regular intervals, and optimal heparin concentration required to prevent gel formation in various media was determined. The biological activity of HPL and FBS was compared by evaluating the growth performance of Vero and Hep-2 cell lines.
Results: Result of platelet count assay revealed the efficiency of HPL preparation process. Growth factor concentrations in HPL were significantly higher than those in FBS, while the protein content of HPL was lower than that of FBS. Stability study data showed that the prepared HPL was stable for up to 15 months at −20C. Ideal heparin concentration to be used in different media was dependent on calcium concentration. Results of cell viability assay showed that HPL was superior to FBS in supporting the growth and proliferation of Vero and Hep-2 cells.
Conclusion: The HPL prepared by the mechanical activation of platelets may serve as an efficient alternative to FBS in cell culture process.
Keywords: Vero cell line, Hep-2 cell line, Heparin, Human platelet lysate, Growth factors, Viability assay


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