Proliferative Capacities and Differentiation Potentials of Human Periodontal Ligament Stem Cells after Slow-freezing Cryopreservation

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Published: May 31, 2022
Keywords:
Cell proliferation, Cryoprotectant agents, Multipotency, Periodontal ligament stem cells, Slow-freezing cryopreservation

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Dea Ajeng Pravita Suendi
Graduate School of Biotechnology, Postgraduate Program, University of Jember, Jember 68121, East Java, Indonesia
Banun Kusumawardani
Graduate School of Biotechnology, Postgraduate Program, University of Jember, Jember 68121, East Java, Indonesia

Abstract

Introduction: The cryopreservation of periodontal ligament stem cells (PDLSCs) required a good combination of CPA composition as a step in the preparation of PDLSCs. This study aimed to analyze the proliferative capacities and differentiation potentials of PDLSCs after slow-freezing cryopreservation with CPA in different combinations. Methods: The fourth passage of the primary PDL cells were examined their fibroblast-like morphology and colony forming unit-fibroblast (CFU-F), and characterized by surface markers for mesenchymal stem cells using flow cytom- etry. PDLSCs were divided into two groups of freshly-PDLSCs (fPDLSCs) and cryopreserved-PDLSCs (cPDLSCs). The PDLSCs were cryopreserved using slow freezing method with CPA in different combinations: 1) 90%FBS+10%D- MEM (FD-group), 2) 90%DMEM+10%DMSO (DDs-group), 3) 90%FBS+10%DMSO (FDs-group), and 4) 100% Cell Banker (CB-group) as positive control. The proliferation of fPDLSCs and cPDLSCs were evaluated by trypan blue dye exclusion method. The multipotency of cells was assessed by Oil Red O, Alizarin Red, and Alcian Blue staining. Re- sults: The primary PDL cells had fibroblast-like morphology and CFU-F ability. They expressed more than 95% posi- tive MSC surface markers of CD90, CD73, CD150, and CD44, but showed less than 2% hematopoietic cell markers of CD11b/CD19/CD34/CD45 and HLA-DR. The cPDLSCs viability of FDs-group was 81.5% and 80% in -80oC and LN2, respectively. The fPDLSCs and cPDLSCs proliferation and doubling time were no statistically significant differ- ence (p>0.05). They could differentiate into adipogenic, osteogenic, and chondrogenic differentiation. Conclusion: The cPDLSCs could maintain their proliferative capacities and differentiation potentials after slow-freezing cryopres- ervation with 90%FBS+10%DMSO in -80oC.

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How to Cite
Suendi, D. A. P., & Kusumawardani, B. (2022). Proliferative Capacities and Differentiation Potentials of Human Periodontal Ligament Stem Cells after Slow-freezing Cryopreservation. Malaysian Journal of Medicine and Health Sciences, 18(3), 125–132. Retrieved from http://mjmhsojs.upm.edu.my/index.php/mjmhs/article/view/369
Issue
Vol. 18 No. 3 (2022)
Section
Original Articles

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