Cloning of human cord blood-mesenchymal stem cells for isolation of enriched cell population of higher proliferation and differentiation potential

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Demerdash Z.; El Baz H.; Ali N.; Mahmoud F.; Mohamed S.; Khalifa R.; Hassan M.; Shawky S.

Journal: Molecular Biology Reports

Publisher: Springer

Publication Date: 11 May 2020

Volume / Issue: Volume 47 / Issue 5

Pages: 3963–3972

ISSN: 3014851

DOI: 10.1007/s11033-020-05489-1

Scopus: View on Scopus

PubMed: 32394306

Document Type: Article

Authors and Affiliations

Demerdash Z., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; El Baz H., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; Ali N., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; Mahmoud F., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; Mohamed S., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; Khalifa R., Clinical and Chemical Pathology Department, Kasr Al-Ainy, Faculty of Medicine, Cairo University, Cairo, Egypt; Hassan M., Immunology Department, Theodor Bilharz Research Institute, Warrak El-Hadar, Giza, 12411, Egypt; Shawky S., Clinical and Chemical Pathology Department, Kasr Al-Ainy, Faculty of Medicine, Cairo University, Cairo, Egypt

Abstract

Heterogeneity of Mesenchymal stem cells (MSCs) imposes limitations for their in vitro expansion and accounts for the lack of reproducibility in some clinical studies. So, this study was designed to isolate and enrich clones of multipotent and self-renewing MSCs from cord blood (CB). Enriched clones with higher proliferation and differentiation potential provide regenerative cells suitable for various clinical demands. MSCA and MSCB original (progenitor) cells were isolated from CB samples, and single cells were cloned by limiting dilution method, in mouse embryonic fibroblast conditioned media. Original MSCs and their single-cell derived clones were characterized by identifying their proliferation rate, immunophenotyping of surface antigens, expression of pluripotency and proliferation genes (Oct4, Sox2, Nanog, KLF4, c-Myc, and PDGFRA), and differentiation potential into multiple lineages (osteogenic, adipogenic, and chondrogenic). Some single-cell clones of MSCA showed a higher proliferation rate and greater differentiation potential than their original cells. However, original MSCB cells were of greater proliferation and differentiation potential than their derived single-cell clones, except for one clone which had comparable results. Cloning of MSCs was attainable when cultured in mouse embryonic fibroblast conditioned media. Single clones with higher proliferation and differentiation potential than their original progenitor cells were obtained by cloning of poorly functioning MSCs progenitor cells, enabling the selection of more therapeutically efficacious MSCs with better performance in clinical applications. Moreover, this study draws attention to the importance of CD105 as a possible MSCs biomarker associated with the multilineage commitment of MSCs. © 2020, Springer Nature B.V.

Keywords

Cloning; Cord blood-mesenchymal stem cells; Differentiation potential; Limiting dilution method; Pluripotency genes; Adipogenesis; Cell Differentiation; Cell Proliferation; Cells, Cultured; Chondrogenesis; Cloning, Molecular; Fetal Blood; Humans; Immunophenotyping; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Reproducibility of Results; Umbilical Cord; endoglin; kruppel like factor 4; membrane antigen; Myc protein; octamer transcription factor 4; platelet derived growth factor alpha receptor; transcription factor NANOG; transcription factor Sox2; Article; bone development; cell cloning; cell culture; cell function; cell isolation; cell population; cell regeneration; cell self-renewal; gene expression; human; human cell; mesenchymal stem cell; umbilical cord blood; cytology; fetus blood; metabolism; molecular cloning; physiology; procedures; reproducibility

Citation Information

Scopus Citations: 7

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