• International Journal of Oral Biology
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• 제37권4호
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• pp.175-180
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• 2012

Skin-derived precursor cells (SKPs) are multipotent, sphere-forming and embryonic neural crest-related precursor cells that can be isolated from dermis. It is known that the properties of porcine SKPs can be enhanced by leukemia inhibitory factor (LIF) which is an essential factor for the generation of embryonic stem cells in mice. In our present study, to enhance or maintain the properties of murine SKPs, LIF was added to the culture medium. SKPs were treated with 1,000 IU LIF for 72 hours after passage 3. Quantitative real time RT-PCR was then performed to quantify the expression of the pluripotent stem cell specific genes Oct4, Nanog, Klf4 and c-Myc, and the neural crest specific genes Snai2 and Ngfr. The results show that the expression of Oct4 is increased in murine SKPs by LIF treatment whereas the level of Ngfr is decreased under these conditions. Interestingly, LIF treatment reduced Nanog expression which is also important for cell proliferation in adult stem cells and for osteogenic induction in mesenchymal stem cells. These findings implicate LIF in the maintenance of stemness in SKPs through the suppression of lineage differentiation and in part through the control of cell proliferation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권6호
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• pp.588-593
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• 2008

There are increasing reports regarding regeneration of the defected tissues using tissue engineering technique. In this technique, multipotential stem cells are essential. There are many potential sources of adult stem cells, such as bone marrow, umbilical cord blood, fat, muscle, dental tissues and skin. Among them, skin is highly accessible and easily obtained with a minimum of donor site complications. Moreover, skin is an abundant adult stem cell sources and has the potential for self-replication and immune privilege. In this study, we isolated skin-derived precursor cells (SKPs) from the ear of adult miniature pigs. In these SKPs, the expression of transcriptional factors, Oct-4, Sox-2, and Nanog were detected by RT-PCR. In vitro osteogenesis and adipogenesis were observed at 3 weeks after transdifferentiations as assayed by positive von Kossa and Oil-red O staining, respectively. In addition, expression of osteocalcin and osteonectin in the osteogenic differentiation medium and $PPAR{\gamma}2$ and aP2 in the adipogenic differentiation medium were detected by RT-PCR. In vitro neurogenesis of porcine SKPs was observed during 24 and 72 hours after treatment of neurogenic differentiation medium. The results of this study suggest that SKPs demonstrate the properties of pluripotence or multipotence and multi-lineage differentiation. This indicates that autogenous SKPs are a reliable and useful source of adult stem cells for regenerative medicine.

• International Journal of Oral Biology
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• 제41권2호
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• pp.69-74
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• 2016

Skin-derived precursors (SKPs) have potential to differentiate to various cell types including osteoblasts, adipocytes and neurons. SKPs are a candidate for cell-based therapy since they are easily accessible and have multipotency. Most mammalian cells are exposed to a low oxygen environment with 1 to 5% $O_2$ concentration in vivo, while 21% $O_2$ concentration is common in in vitro culture. The difference between in vitro and in vivo $O_2$ concentration may affect to the behavior of cultured cells. In this report, we investigated the effect of hypoxic condition on stemness and proliferation of SKPs. The results indicated that SKPs exposed to hypoxic condition for 5 days showed no change in proliferation. In terms of mRNA expression, hypoxia maintained expression of stemness markers; whereas, oncogenes, such as Klf4 and c-Myc, were downregulated, and the expression of Nestin, related to cancer migration, was also downregulated. Thus, SKPs cultured in hypoxia may reduce the risk of cancer in SKP cell-based therapy.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권3호
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• pp.198-206
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• 2009

Purpose: The aims of this study were to assess the in vitro co-culturing pattern of isolated skin-derived precursor cells (SKPs) with a mixed demineralized bone (DMB) and fibrin glue scaffold and to evaluate in vivo osteogenesis after transplantation of autogenous SKPs with a these mixed scaffold in the animal's mandibular defects. Materials and Methods: We isolated SKPs from the ears of adult 4 miniature pigs. The isolated SKPs were co-cultured with a mixed DMB and fibrin glue scaffold in a non-osteogenic medium for 1, 2, and 4 weeks. Histological characteristics of in vitro co-cultured cells and scaffold were evaluated. $1{\times}10^7\;cells/100\;{\mu}l$ of autogenous porcine SKPs were grafted into the mandibular defects with a DMB and fibrin glue scaffold. In the control sites, only a scaffold was grafted, without SKPs. After two animals each were euthanized at 2 and 4 weeks after grafting, the in vivo osteogenesis was evaluated with histolomorphometric and osteocalcin immunohistochemical studies. Results: Homogeneously shaped skin-derived cells were isolated from porcine ear skin after 3 or 4 weeks of primary culture. In vitro osteogenic differentiation of SKPs was observed after co-culturing with a DMB and fibrin glue scaffold in a non-osteogenic medium. Von Kossa-positive bone minerals were also noted in the co-cultured medium at 4 weeks. As the culture time progressed, the number of observable cells increased. Trabecular new bone formation and osteocalcin expression were more pronounced in the SKP-grafted group compared to the control group. Conclusion: These findings suggest that autogenous SKP grafting with a DMB and fibrin glue scaffold can serve as a useful alternative to bone grafting technique.

• 한국응용과학기술학회지
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• 제13권3호
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• pp.15-24
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• 1996

Essentiality was proposed in the field of lipid by Burr and Burr in 1929. When rats were raised on the fat-free diet, their growth retarded and their skin and tails showed the characteristic deficient symptoms, which were relieved by the addition of ${\omega}6(n-6)$ polyunsaturated fatty acids as linoleic(LA) and arachidonic(AA) acids to the basal diet. LA is dehydrogenated to ${\gamma}-linolenic$ acid(GLNA) by ${\Delta}6$ desaturase, then GLNA is 2 carbon chain elongated by elongase to $dihomo-{\gamma}-linolenic$ acid(DGLNA), which is desaturated by ${\Delta}5$ desaturase to AA. These acids are called LA family or ${\omega}6(n-6)$ polyunsaturated fatty acids(PUFA). ${\alpha}-Linolenic$ acid(ALNA) is converted through the series of desaturation and elongation steps to docosahexaenic acid(DHA) via eicosapentaenoic acid(EPA). These acids belong to ALNA family or ${\omega}3(n-3)$PUFA. Human who consume large amounts of EPA and DHA, which are present in fatty fish and fish oils, have increased levels of these two fatty acids in their plasma and tissue lipids at the expense of LA and AA. Alternately, vegetarians, whose intake of LA in high, have more elevated levels of LA and AA and lower levels of EPA and DHA in plasma lipids and in cell membranes than omnivores. AA and EPA are metabolized to substances called eicosanoids. Those derived form AA are known as prostanocids(prostaglandins and prostacyclins) of the 2-types and leukotrienes of the 4-series, whereas those derived from EPA are known as prostanoids of the 3-types and leukotrienes of the 5-series. DGLNA is a precursor of the 1-types of prostaglandins. The metabolites of AA and EPA have competitive functions. Ingestion of EPA from fish or fish oil replaces AA from membrane phospholipids in practically all cells. So this leads to a more physiological state characterized by the production of proatanoids and leukotrienes that have antithrombic, antichemotactic, antivasoconstrictive and antiinflammatory properties. It is evident that ${\omega}3$ fatty acids can affect a number of chronic diseases through eicosanoids alone.