Characterization of Human Gingiva- and PDL-derived Progenitor Cells in Xeno-free Conditions

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Characterization of Human Gingiva- and PDL-derived Progenitor Cells in Xeno-free Conditions

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Publication Conference Poster
Title Characterization of Human Gingiva- and PDL-derived Progenitor Cells in Xeno-free Conditions
Author Shanbhag, Siddharth ; Ahmed, Samih Mohamed ; Suliman, Salwa ; Stavropoulos, Andreas ; Bolstad, Anne Isine ; Mustafa, Kamal
Date 2018
English abstract
Objectives: Periodontal tissues represent a clinically relevant and less invasive source of progenitor cells compared to bone marrow, for periodontal- and/or alveolar bone-tissue engineering (P-/BTE). The safety and efficacy of clinical-grade stem cells can be enhanced via substitution of xenogeneic supplements and three-dimensional (3-D) cell culture, respectively, to simulate the in vivo microenvironment more closely. The objective of this study was to comprehensively characterize progenitor cells derived from human gingiva (GPCs) and periodontal-ligament (PDL; PPCs) in xeno-free conditions for use in P-/BTE. Methods: In preliminary experiments, pooled human platelet lysate (PL) was identified as the optimal xeno-substitute for fetal bovine serum (FBS) in stem cell cultures. Primary GPCs and PPCs were isolated and expanded as monolayers in PL- and FBS-supplemented media; passage 3-5 cells were used in experiments. Bone marrow mesenchymal stem cells (BMSCs) were used as a reference. Growth kinetics were compared via cell proliferation and colony forming-unit (CFU) assays. GPCs and PPCs were characterized via cytometric expression of stromal markers, multi-lineage (osteogenic, adipogenic and chondrogenic) differentiation potential, and secretory cytokine profiles. 3-D sphere cultures of GPCs and PPCs were established, and the expression of stemness- and osteogenesis-related markers in 3-D and monolayer cultures was evaluated via gene expression and immunocytochemistry. Results: GPCs and PPCs in both FBS and PL showed characteristic fibroblastic morphology and stromal phenotype (highly positive for CD105/CD90/CD73 and negative for CD34/CD45/HLA-DR). Cell proliferation and CFU efficiency were superior in PL compared to FBS. GPCs and PPCs expanded in both PL and FBS showed multi-lineage differentiation comparable to BMSCs; notably, osteogenic differentiation was enhanced in GPCs expanded in PL. 3-D spheres of GPCs and PPCs were formed and maintained for up to 7 days in xeno-free suspension culture. Notably, expression of stemness- (Sox2, Oct4, Nanog) and osteogenesis-related markers (Runx2, Osx, BMP2) was significantly upregulated in GPC- and PPC-derived spheres vs. monolayers. Conclusions: PPCs and particularly GPCs, due to easy access, cultured in 3-D xeno-free conditions represents a promising strategy for P-/BTE. Student Presenter This abstract is based on research that was funded entirely or partially by an outside source: International Team for Implantology (117/2015), Helse Vest Norway (912048) Disclosure Statement: The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE
Conference
2018 IADR/PER General Session & Exhibition Pan European Regional Congress (July 25-28, 2018 : London, England)
Language eng (iso)
Subject Medicine
Research Subject Categories::ODONTOLOGY
Note The 96th General Session of the International Association for Dental R... (see Details for more)
Handle http://hdl.handle.net/2043/27135 Permalink to this page
Link https://iadr2018.zerista.com/... (external link to related web page)
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