Passaging of CB-MSCs at high and low cell densities: Impact on telomere length and differentiation potentials

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Elbaz H.; Demerdash Z.; Elzallat M.; Mahmoud F.S.; Hassan S.; Hassan M.; Taha T.; Rasheed D.M.; Gaafar T.

Journal: Gazzetta Medica Italiana Archivio per le Scienze Mediche

Publisher: Edizioni Minerva Medica

Publication Date: February 2022

Volume / Issue: Volume 180 / Issue 10

Pages: 530–538

ISSN: 3933660

DOI: 10.23736/S0393-3660.19.04277-3

Scopus: View on Scopus

Document Type: Article

Authors and Affiliations

Elbaz H., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Demerdash Z., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Elzallat M., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Mahmoud F.S., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Hassan S., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Hassan M., Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt; Taha T., Department of Reproductive Health Research, National Research Center, Giza, Egypt; Rasheed D.M., Department of Clinical and Chemical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt; Gaafar T., Department of Clinical and Chemical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt

Abstract

BACKGROUND: Mesenchymal stem cells (MSCs) are considered valuable candidates for cell-based therapy. Identifying factors influencing MSCs stemness properties, namely self-renewal, differentiation, and proliferation potential, could have a great impact on their therapeutic applications. This study was designed to define the favorable cell density with a positive impact on the stemness properties of MSCs. METHODS: MSCs were isolated from cord blood and cultured at different seeding densities till reaching senescence. MSCs were subjected to population doubling (PD) rate calculation, differentiation into osteogenic, adipogenic, and chondrogenic lineages, pluripotency genes expression analysis and absolute telomere length measurement. RESULTS: MSCs showed better morphology at low seeding density than at high-seeding density. Moreover, the PDof MSCs was higher at low seeding densities than at high-seeding densities. However, both differentiation potential of MSCs and their telomere length were lower at low-seeding densities than at high-seeding densities. Regarding pluripotency gene expression, cells cultured at high-cell density showed the highest expression of Sox2, Nanog, and c-Myc; while those cultured at lower-cell densities showed the highest expression of Oct4, Klf4, and PDGFRa. CONCLUSIONS: Choosing the best seeding density is crucial for optimizing the clinical application effectiveness of MSCs, as the low seeding density has unfavorable effects on their telomere length and differentiation potentials. © 2021 Edizioni Minerva Medica. All rights reserved.

Keywords

Cell differentiation; Mesenchymal stem cells; Population; Telomere; 5' nucleotidase; alizarin; CD34 antigen; endoglin; glutamine; kruppel like factor 4; octamer transcription factor 4; phycoerythrin; platelet derived growth factor alpha receptor; receptor type tyrosine protein phosphatase C; streptomycin; Thy 1 membrane glycoprotein; transcription factor NANOG; transcription factor Sox2; trypsin; Article; cell culture; cell density; cell proliferation; chondrocyte; colony formation; controlled study; female; fetal bovine serum; gene expression; gene expression profiling; genomic instability; human; human cell; immunophenotyping; karyotyping; mesenchymal stem cell; mononuclear cell; oncogene c myc; osteoblast; peripheral blood mononuclear cell; pluripotent stem cell; protein expression; real time polymerase chain reaction; regenerative medicine; reverse transcription polymerase chain reaction; RNA extraction; senescence; telomere length; umbilical cord blood

Citation Information

Scopus Citations: 0

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