Improvement and Comparison of Mesenchymal Stem Cell Cultivation from Umbilical Cord Using Different Concentrations of Fetal Bovine Serum and Platelets Lysate

Abstract

Abstract Introduction In recent years there seems to be an unbounded interest concerning MSCs, this is mainly attributed to their exciting characteristics including long-term ex vivo proliferation, multilineage potential and immunomodulatory properties. In this regard MSCs emerge as attractive candidates for various therapeutic applications especially for the treatment of a range of diseases that are not curable by current therapies. MSCs obtained from the Wharton's Jelly (WJ) and umbilical cords blood (UCB) have gained much attention over the last years since they can be easily isolated, without painful isolation procedure and without any ethical concerns, from a tissue which is discarded after birth. Furthermore umbilical cord-derived MSCs represent a more primitive population than their adult counterparts, opening new perspectives for cell-based therapies. In this study we aimed to establish xenogenic-free expansion protocol to be compliant with good manufacturing practice (GMP) guidelines. We used human platelets lysate (HPL) as alternative to fetal bovine serum which used in most protocol of MSCs expansion, since platelet-derived growth factors can promote MSC ex-vivo expansion. Materials and Methods Nine Cord Blood samples and Ten Wharton′s Jelly samples were used for MSCs isolation and one Bone Marrow sample was used as control. Platelets concentrate obtained from the blood bank were activated by calcium gluconate to get platelets lysate. Results Our study showed that the expansion potential of HPL medium cultured MSCs was significantly higher than FBS medium, and the HPL is able to decrease the time required to reach confluence, and to increase Colony Forming Unit Fibroblast size and count of viable cells as compared to the FBS medium. Furthermore, the HPL exhibited dose dependent proliferation rate and our results display that the 10% HPL is most effective concentration in term of culture consideration and cells characterization. In addition the results of immunophenotyping assay showed that the HPL cultured MSCs expressed higher concentration of CD44 and CD73 with high flow rate compared to those cultured in FBS media. Conclusions The comparative analysis of the MSCs from the Two sources in different concentration of HPL and FBS indicated that the W.J is more promising source for MSCs compared to C.B, and the HPL is the safe, powerful and cost effective supplement for expansion of human MSCs.