Isolation and identification of human periodontal ligament stem cells in vitro.
- Author:
Tao SHEN
1
;
Hui-jun CHANG
;
Cong-xiang JIAN
;
Yan-chun YANG
;
Ji-xiang ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Cell Differentiation; Cell Separation; Humans; In Vitro Techniques; Osteoblasts; Periodontal Ligament; Stem Cells
- From: West China Journal of Stomatology 2011;29(1):71-78
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo isolate and identify human periodontal ligament stem cells (PDLSC) by improved methods and assess the characteristics of PDLSC ex vivo.
METHODSThe periodontal ligament cells were obtained from the healthy impacted third molars and teeth extracted for orthodontic purposes and used to isolate PDLSC by limiting dilution assay. PDLSC were cultured and expanded in alpha-MEM supplemented with 10% FBS. Colony-forming assay, immunohistochemistry, flow cytometry, osteogenic and adipogenic induction were used to identify PDLSC.
RESULTSThe obtained cells had high colony-forming efficiency and were positive staining for vimentin and negative for pancytokeratin. Flow cytometry revealed that the isolated cells were positive for STRO-1 and CD146 antibodies and most were in the G0/G1 phase of cell cycle. Under specific conditions, they could differentiate to the osteoblast and adipocyte lineages in vitro.
CONCLUSIONLimiting dilution assay is an effective method to isolate PDLSC and the single-cell-derived colonies demonstrate the properties of stem cells in vitro.
