Assessment of an efficient xeno-free culture system of human periodontal ligament stem cells.
The possibility of transplanting adult stem cells into damaged organs has opened new prospects for the treatment of several human pathologies. The purpose of this study was to develop a culture system for the expansion and production of human Periodontal Ligament Stem Cells (hPDLSCs) using a new xeno-free media formulation and ensuring the maintenance of the stem cells features comprising the multiple passage expansion, mesengenic lineage differentiation, cellular phenotype, and genomic stability, essential elements for conforming to translation to cell therapy. Somatic stem cells were isolated from the human periodontium using a minimally invasive periodontal access flap surgery in healthy donors. Expanded hPDLSCs in a xeno-free culture showed the morphological features of stem cells, expressed the markers associated with pluripotency, and a normal karyotype. Under appropriate culture conditions, hPDLSCs presented adipogenic and osteogenic potential; indeed, a very high accumulation of lipid droplets was evident in the cytoplasm of adipogenic-induced cells, and indisputable evidence of osteogenic differentiation, investigated by transmission electron microscopy, and analyzed for gene expression analysis has been shown. Based on these data, the novel xeno-free culture method might provide the basis for Good Manufacturing Procedure culture of autologous stem cells, readily accessible from human periodontium, and can be a resource to facilitate their use in human clinical studies for potential therapeutic regeneration.
1 Laboratory of Stem Cells and Regenerative Medicine, Department of Medical, Oral and Biotechnological Sciences, School of Medicine and Health Sciences, University "G. d'Annunzio" Chieti-Pescara , Chieti, Italy .
Oriana
O
Trubiani
Adriano
A
Piattelli
Valentina
V
Gatta
Marco
M
Marchisio
Francesca
F
Diomede
Marco
M
D'Aurora
Ilaria
I
Merciaro
Laura
L
Pierdomenico
Nadir Mario
NM
Maraldi
Nicoletta
N
Zini
D000328:Adult; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D016130:Immunophenotyping; D007621:Karyotyping; D039902:Multipotent Stem Cells / Q000166:cytology; D010012:Osteogenesis / Q000235:genetics; D010513:Periodontal Ligament / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D055815:Young Adult