The Effect of Platelet Lysate on Expansion and Differentiation Megakaryocyte Progenitor Cells from Cord Blood CD34+ enriched Cells

Yoda Yaghoubi; Ali  Hassanzadeh; Adel  Naimi; Sepehr  Abdolahi; Mehdi  Yousefi; Leili  Aghebati-Maleki; Majid  Zamani

doi:10.18502/ijpho.v11i3.6563

Yoda Yaghoubi Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
Ali Hassanzadeh Department of Tissue Engineering and Applied Cell Sciences, Tehran University of Medical Sciences, Tehran, Iran
Adel Naimi Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran
Sepehr Abdolahi Department of Biology, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran
Mehdi Yousefi Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Leili Aghebati-Maleki Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Majid Zamani Department of Medical Laboratory Sciences, Faculty of Allied Medicine, Gonabad University of Medical Sciences, Gonabad, Iran

DOI: https://doi.org/10.18502/ijpho.v11i3.6563

Keywords: Cord Blood Stem Cell Transplantation; Hematopoietic Stem Cells; Megakaryocyte Progenitor Cells; Platelet-Derived Growth Factor

Abstract

Background: Umbilical cord blood hematopoietic stem cells (UCB-HSCs) are an attractive source for transplantation. The generation of megakaryocyte-committed cells could lead to shorten period of thrombocytopenia after HSCs transplantation. Platelet lysate (PL) unlike fetal bovine serum (FBS) can prevent immune problems as well as avert transmission of certain diseases to the recipient. In this study, the authors aimed to assess the effect of PL on UCB CD34⁺ cells expansion and megakaryocyte differentiation.

Materials and Methods: In this experimental study, PL prepared and the subsequent isolation of UCB CD34⁺ cells were done by magnetic cell sorting. The isolated cells were cultivated in Iscove’s Modified Dulbecco’s medium (IMDM) supplemented with PL or FBS. Cell expansion was evaluated using Trypan blue. Furthermore, Flow cytometry using monoclonal antibodies (CD41-FITC and CD42b-PE) and the expression of specific genes including GATA1, GATA2, FLI1, NFE2, and RUNX1 via real-time PCR were performed to evaluate the megakaryocyte differentiation.

Results: The results showed that PL insignificantly enhanced UCB CD34⁺ cell expansion (32.83± 8.47 fold in FBS and 41.67± 10.31 fold in PL containing media). Besides, flow cytometry results showed that expression of CD41 was increased markedly (37.81± 4.78 fold in FBS and 45.78 ± 7.37 in PL containing media, P-value <0.05) but the elevation of CD42b (10.53 ± 2.13 and 13.20 ± 2.06 in FBS and PL containing media, respectively) was not significant (P-value = 0.051). The results of real-time PCR demonstrated a notable increase in GATA binding protein 1 (1.58, P-value <0.01), GATA binding protein 2 (2.45, P-value <0.001), RUNX family transcription factor 1 (1.60, P-value <0.01), Fli-1 proto-oncogene (1.87, P-value <0.001) in PL supplemented media, however, the increase of Nuclear Factor-Erythroid 2 gene expression was not significant in PL supplemented media (P-value = 0.11).

Conclusion: PL improved UCB CD34⁺ cells expansion and megakaryocyte differentiation compared to FBS.