Culture and Characterization of Mesenchymal Stem Cells From Human Gingival Tissue

TLDR

Mesenchymal stem cells (MSCs) offer high proliferation and differentiation potential, making them attractive for tissue engineering, yet identifying MSCs capable of regenerating the periodontium remains challenging due to the lack of predictable regeneration procedures. The study aimed to identify mesenchymal stem cells in human gingival connective tissue and characterize their morphology and functional properties. Gingival connective tissue from five healthy donors was processed, cultured for 3–4 weeks, and then characterized by flow cytometry, lineage differentiation assays, and immunoregulation testing. The cultured gingival cells met MSC criteria, expressed CD90, CD105, CD73, CD44, and CD13 while lacking hematopoietic markers, differentiated into adipogenic, chondrogenic, and osteogenic lineages, and exhibited immunomodulatory activity, indicating that gingival tissue is a viable MSC source for regenerative applications.

Abstract

Tissue engineering using mesenchymal stem cells (MSCs) is a recent therapeutic modality that has several advantages. MSCs have high proliferation potential and may be manipulated to permit differentiation before being transplanted, suggesting they may be an ideal candidate for regenerative procedures. Precise identification of cells capable of regenerating the periodontium is valuable because no predictable regeneration procedure has yet been described. The purpose of this study is to determine the presence of MSCs in human gingival connective tissue and their morphologic and functional characteristics.Gingival connective tissue samples were obtained from five healthy students. The samples were deepithelialized, leaving only connective tissue. The explants were minced and cultured on tissue culture dishes for 3 to 4 weeks, after which cells were characterized by flow cytometry. Differentiation into osteogenic, chondrogenic, and adipogenic lineages was induced and evaluated by culture staining. An immunoregulation assay was also performed.The results show that gingival tissue cells fulfill the minimal criteria proposed by the International Society for Cellular Therapy to be defined as MSCs. Cell characterization was consistently positive for CD90, CD105, CD73, CD44, and CD13 markers and negative for hematopoietic markers CD34, CD38, CD45, and CD54. We observed differentiation in positive staining of adipogenic, chondrogenic, and osteogenic lineages. Furthermore, gingival cells showed immunomodulative capacity.Gingival connective tissue could be a reservoir of MSCs that could be used in regenerative procedures based on tissue engineering.