Crosstalk on Cell Behavior in Interactive Cocultures of hMSCs with Various Oral Cell Types

Abstract

When prospectively applied for regenerative therapies, human bone-marrow-derived mesenchymal stem cells (hMSCs) interact with the locally residing host cells. With respect to the developmentally particular origin of oral cells, little is known about the putatively discriminative behavioral responses of hMSCs in interaction with various oral cell types, including human alveolar bone osteoblasts (hOAs), periodontal ligament fibroblasts (hPDLs), and gingival fibroblasts (hGFs). To assess the crosstalk between hMSCs and oral cells, interactive cocultures were established by combining well-characterized hMSCs with hOAs, hPDLs, or hGFs, and the behavioral hMSC aspects, that is, proliferation and gene expression, were measured by employing a 5-bromo-2'-deoxyuridine assay and real-time polymerase chain reaction, while apoptosis was quantified by in situ cell death detection kit. hMSCs expressed the typical antigen spectrum lacking CD34, CD45, CD14, CD19, and HLA-DR, while expressing CD73, CD90, and CD105, and could successfully be transformed into adipocytes, osteocytes, and chondrocytes. Monocultured control hMSCs proliferated readily, whereas a general reduction of BrdU-labeled cells was observed in cocultures. Globally, upon extending time periods, interactive coculture combinations of hMSCs with hOAs reduced both osteogenic gene and stem cell marker transcription in hMSCs, a phenomenon appearing less pronounced by combining hMSCs with hPDLs, such that the observed effects in terms of proliferation and gene expression followed the same ranking: hOAs>hGFs>hPDLs. Vice versa, in interactive hMSC cocultures, the cell survival rate was significantly increased, irrespective from the combined coculture cell counterpart. Our results show for the first time that behavior of hMSCs reflected by proliferation and gene expression was governed by interaction with various oral cells in a cell-type-discriminative manner. In addition, hMSC coculture restrains apoptosis, such that influences on cell behavior appear as a crosstalk. In summary, interactive cocultures render the basis for a prospective prediction of mutual cell behavior in hMSC-based oral tissue regeneration disclosing that oral cells shift hMSC behavior from proliferation to differentiation and apoptosis-repressing features.

Assessment of an Efficient Xeno-Free Culture System of Human Periodontal Ligament Stem Cells

“…58 Then, hPDLSCs can be manipulated ex vivo under xeno-free conditions without compromising their functional properties and offering various potential advantages when compared with a standard culture system; these results are according to current literature showing that the modified serum-free media support the expansion of oral stem cells and the maintenance of their multipotency. 21 …”

Cell-cell interaction between vocal fold fibroblasts and bone marrow mesenchymal stromal cells in three-dimensional hyaluronan hydrogel

“…The literature supports the attenuation of proliferation in cardiac fibroblasts treated with BM‐MSCs conditioned media (Ohnishi et al ., ) and of ferret vocal fold scar fibroblasts co‐cultured with adipose‐derived stromal cells (Kumai et al ., ). Conversely, decreased viability of MSCs has been measured when BM‐MSCs were co‐cultured with gingival fibroblasts (Proksch et al ., ). Given that all cell types demonstrated decreased proliferation, there appears to be cross‐talk in 3D co‐culture between BM‐MSCs and VFFs.…”

Mesenchymal Stem Cells and Periodontal Regeneration

“…In these cases, it is postulated that transplanted MSCs are acting principally through mechanisms involving the paracrine regulation of immune responses and by release of trophic cytokines that direct endogenous cell regeneration. Additionally, the possibility of crosstalk between implanted MSCs and local cells has been described in vitro [64].…”