In VitroandIn VivoOsteogenesis of Porcine Skin-Derived Mesenchymal Stem Cell–like Cells with a Demineralized Bone and Fibrin Glue Scaffold

Abstract

In vitro and in vivo osteogenesis of skin-derived mesenchymal stem cell-like cells (SDMSCs) with a demineralized bone (DMB) and fibrin glue scaffold were compared. SDMSCs isolated from the ears of adult miniature pigs were evaluated for the expression of transcriptional factors (Oct-4, Sox-2, and Nanog) and MSC marker proteins (CD29, CD44, CD90, and vimentin). The isolated SDMSCs were cocultured in vitro with a mixed DMB and fibrin glue scaffold in a nonosteogenic medium for 1, 2, and 4 weeks. Osteonectin, osteocalcin, and Runx2 were expressed during the culture period and reached maximum at 2 weeks after in vitro coculture. von Kossa-positive bone minerals were also noted in the cocultured medium at 4 weeks. Autogenous porcine SDMSCs (1 x 10(7)) labeled with a tracking dye, PKH26, were grafted into the maxillary sinus with a DMB and fibrin glue scaffold. In the contralateral side, only a scaffold was grafted without SDMSCs (control). In vivo osteogenesis was evaluated from two animals euthanized at 2 and 4 weeks after grafting. In vivo PKH26 staining was detected in all the specimens at both time points. Trabecular bone formation and osteocalcin expression were more pronounced around the grafted materials in the SDMSC-grafted group compared with the control group. New bone generation was initiated from the periphery to the center of the grafted material. The number of proliferating cells increased over time and reached a peak at 4 weeks in both in vivo and in vitro specimens. These findings suggest that autogenous SDMSC grafting with a DMB and fibrin glue scaffold can serve as a predictable alternative to bone grafting in the maxillary sinus floor.

Isolation of human mesenchymal stem cells from the skin and their neurogenic differentiationin vitro

“…Similarly, in this study, human skin-derived cells were isolated from the submandibular skin segments, and multipotent MSC characteristics were detected, i.e., expression of transcription factors (Oct 4 and Nanog), detection of MSC markers (CD29, CD44, CD90, CD105, and vimentin), and potential of in vitro osteogenic, adipogenic, and chondrogenic differentiations. Interestingly, unlike the porcine SMSCs of a previous study 15 , we could not observe Sox 2 expression in the hSMSCs of this study. This is consistent with the result of other research, i.e., high visibility of Oct 4 and Nanog but negative expression of Sox 2 in human BMSCs 24 .…”

“…A) All experiments were authorized by the Gyeongsang National University Hospital Ethics Committee, and the patients gave their informed consent to tissue donation. Fresh human skin samples were transported to the laboratory, and hSMSCs were isolated as previous protocol 15 . Briefly, all hairs and subcutaneous fat tissues were removed, and the samples were then cut into 1-3 mm 2 explants containing the epidermis and dermis.…”

“…Several researchers have demonstrated that there are several different types of stem cells -such as skin-derived precursors (SKPs), skin-derived mesenchymal stem cells (SMSCs), and epidermal stem cells --in the dermis and epidermis of skins [10][11][12][13][14] . In the previous study, we isolated porcine skin-derived cells from the ear skin of miniature pigs and showed the multipotency and MSC characteristics 15 .…”

Analysis of Oocyte-Like Cells Differentiated from Porcine Fetal Skin-Derived Stem Cells

“…Previous works have reported the characterization and differentiation of porcine somatic stem cells [1][2][3][4][5][6][7][8][9][10]. The generation of oocytes from somatic stem cells in vitro may provide a valuable model for identifying factors involved in germ cell formation and oocyte differentiation.…”

Nonadherent culture method downregulates stem cell antigen-1 expression in mouse bone marrow mesenchymal stem cells

“…These cells can be isolated from multiple types of tissue, including bone marrow (BM), skin, adipose and umbilical cord tissue (1)(2)(3), and are able to differentiate into osteoblasts, adipocytes, chondrocytes and myocardial cells in vitro (4,5). Previous studies have confirmed that MSCs are also able to secrete bioactive factors that alter the milieu of dysfunctional tissues (6,7).…”