Localization and characterization of human palatal periosteum stem cells in serum‐free, xeno‐free medium for clinical use

Abstract

Harvesting, expanding, and re‐implanting osteogenic mesenchymal stem cells (MSCs) avoids the donor‐site morbidity associated with autogenous grafting from bone marrow. Mesenchymal stem cells sourced from the palatal periosteum could be an alternative to isolation of such cells using bone marrow aspiration procedures. For safe use in human therapy, MSCs should be expanded in culture medium that is free from animal or human‐derived serum. In this study we localized, quantified, and characterized MSCs from palatal periosteum cultured in serum‐free, xeno‐free Essential 8 medium. A portion of the palatal periosteum tissues from three patients were dual‐immunostained with MSC‐specific markers (CD105, CD90, and CD73). The remaining portions were expanded in culture, and the isolated MSCs were analyzed using flow cytometry and tri‐lineage differentiation. Palatal periosteum sections were found to contain CD105‐, CD90‐, and CD73‐positive cells. The cultured cells were 73.0 ± 6.7% (mean ± SD) positive for all three MSC‐specific markers and were without hematopoietic stem cell (HSC) markers 0.5 ± 0.3% (mean ± SD). Tri‐lineage differentiation analysis confirmed that palatal periosteum cells could become adipoblasts, chondroblasts, and osteoblasts. The results demonstrate that palatal‐derived MSCs could be detected in situ within small niches, and when expanded in serum‐free, xeno‐free medium represent a viable source of MSCs for clinical use.

Immunocytochemical characterization of primary teeth pulp stem cells from three stages of resorption in serum‐free medium

“…MSCs cultured in serum‐free media and with multilineage differentiation capability have been reported to down‐regulate CD105, SSEA4, CD146 and Stro‐1 expression and that this down‐regulation is more marked with passage number. They, however, have been reported to maintain high levels of CD73 and CD90 expression 11,12,58 . There is emerging evidence that serum‐free culture of primary DPSCs results in down‐regulation of CD105, while retaining their NCSC phenotype and multilineage differentiation capability 59 …”

Optimizing growth media for <italic>in vitro</italic> expansion of human bone marrow-derived mesenchymal stem cells

“…Naung et al demonstrate that palatal-derived MSCs could become adipocytes, chondrocytes, and osteoblasts under the culture of serum-free media and xeno-free media. 47 A comparative study's results showed that Wharton's jelly derived mesenchymal stem cells cultured in serum-free, xeno-free medium exhibit superior growth kinetics and functional angiogenesis, alongside other MSC characteristics. 48 In this study, StemPro supplement (group #6) and Essential 8 supplement (E8s, group #4) belong to serum-free and xeno-free media respectively.…”

“…Although previous studied have tested that serum-free media/xeno-free culture media can augment the growth potential of MSCs. [46][47][48] However, additional studies demonstrated that serum-free media result in reduced the proliferative capacity of MSCs, 53 and cannot promote hMSC growth without the addition of cytokines and/or growth factors, possibly since serum induces intracellular calcium oscillations, which are vital to stem cell proliferation and differentiation. 54 Studies have demonstrated that cytokines and/or growth factors influence the proliferation of MSCs.…”

Spatial Distributions, Characteristics, and Applications of Craniofacial Stem Cells

“…Wang et al reported that vitamin C and vitamin D were ideal stimulants of craniofacial PDLSCs for osteoblast differentiation in vitro [ 26 ]. Naung et al proposed a protocol to cultivate palate periosteum-derived MSCs in serum-free and xeno-free medium, which could become a useful source of MSCs for clinical applications [ 86 ]. Moreover, a recent study found that induced pluripotent stem cells (iPSCs) generated from human jaw periosteum cells expressed MSC markers and possessed strong mineralization ability [ 87 ].…”