IADR Abstract Archives
Differentiation of Bone Marrow-derived Mesenchymal Stem Cells Into Oral Epithelial-like Cells by Tissue-specific Extracellular Matrix
Objectives: Objective: Bone marrow-derived mesenchymal stem cells (BM-MSC) are defined by their ability to self-renew and differentiate into osteoblasts, chondrocytes, adipocytes, and myocytes. In addition to these mesenchymal lineages, BM-MSC differentiation into the epithelial lineage has been reported; however, mechanisms of MSC epithelial differentiation remain poorly understood and the few data available are controversial. Stem cells reside in their niches, mainly composed of extracellular matrix (ECM) proteins, and different exogenous factors. The ECM serves as a structural support for the stem cells providing critical signals that regulate self-renewal, proliferation and differentiation of the stem cells. The goal of our study was to commit BM-MSC to the oral epithelial lineage using tissue-specific ECM, acellular dermal matrices, for the repair of oral epithelial defects.
Methods: 1x106 BM-MSC were seeded onto 1cm2 acellular dermal matrices and cultured submerged in growth medium (aMEM containing 15% FBS) for 7 days, then transferred to an air-liquid interface containing epithelial induction medium [(DMEM and Ham’s F-12 mixed 3:1, supplemented with 5% FBS, insulin (5 mg/ml), hydrocortisone (0.4 mg/ml), epidermal growth factor (10ng/ml), T3 (2 ×10-8 M), adenine (1.8×10-1 M), transferrin (5 mg/ml), cholera toxin (10-7 M)]. After 21 days in culture, the acellular dermal matrices containing cells were fixed in 10% formaldehyde and stained with hematoxylin and eosin (H&E) and observed to examine tissue architecture.
Results: Histological analysis showed BM-MSC forming an adherent monolayer after 7 days in culture. After 21 days in an air-liquid interface, BM-MSC appeared cuboidal in shape forming a multiple cell layers with a higher cell density on top of the basement membrane and rete ridges projecting into the underlying connective tissue.
Conclusions: Acellular dermal matrices provide a tissue-specific niche for supporting BM-MSC to proliferate and form a multiple cell stratified oral epithelial-like structure in vitro.
Methods: 1x106 BM-MSC were seeded onto 1cm2 acellular dermal matrices and cultured submerged in growth medium (aMEM containing 15% FBS) for 7 days, then transferred to an air-liquid interface containing epithelial induction medium [(DMEM and Ham’s F-12 mixed 3:1, supplemented with 5% FBS, insulin (5 mg/ml), hydrocortisone (0.4 mg/ml), epidermal growth factor (10ng/ml), T3 (2 ×10-8 M), adenine (1.8×10-1 M), transferrin (5 mg/ml), cholera toxin (10-7 M)]. After 21 days in culture, the acellular dermal matrices containing cells were fixed in 10% formaldehyde and stained with hematoxylin and eosin (H&E) and observed to examine tissue architecture.
Results: Histological analysis showed BM-MSC forming an adherent monolayer after 7 days in culture. After 21 days in an air-liquid interface, BM-MSC appeared cuboidal in shape forming a multiple cell layers with a higher cell density on top of the basement membrane and rete ridges projecting into the underlying connective tissue.
Conclusions: Acellular dermal matrices provide a tissue-specific niche for supporting BM-MSC to proliferate and form a multiple cell stratified oral epithelial-like structure in vitro.