ISOLATION OF PORCINE MULTIPOTENTIAL SKIN-DERIVED PRECURSOR CELLS AND ITS MULTILINEAGE DIFFERENTIATION

미니돼지에서 다능성 피부유래 전구세포의 추출과 이의 다배엽 세포로의 분화유도에 대한 연구

  • Choi, Moon-Jeong (Department Oral & Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University) ;
  • Byun, June-Ho (Department Oral & Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University) ;
  • Kang, Eun-Ju (College of Veterinary Medicine, Gyeongsang National University) ;
  • Rho, Gyu-Jin (College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Uk-Kyu (Department of Oral & Maxillofacial Surgery, School of Dentistry, Pusan National University) ;
  • Kim, Jong-Ryoul (Department of Oral & Maxillofacial Surgery, School of Dentistry, Pusan National University) ;
  • Park, Bong-Wook (Department Oral & Maxillofacial Surgery, School of Medicine and Institute of Health Science, Gyeongsang National University)
  • 최문정 (경상대학교 의학전문대학원 치과학교실 구강악안면외과, 경상대학교 건강과학연구원) ;
  • 변준호 (경상대학교 의학전문대학원 치과학교실 구강악안면외과, 경상대학교 건강과학연구원호) ;
  • 강은주 (경상대학교 수의과대학 수의산과학) ;
  • 노규진 (경상대학교 수의과대학 수의산과학) ;
  • 김종렬 (부산대학교 치의학전문대학원 구강악안면외과학교실) ;
  • 김욱규 (부산대학교 치의학전문대학원 구강악안면외과학교실) ;
  • 박봉욱 (경상대학교 의학전문대학원 치과학교실 구강악안면외과, 경상대학교 건강과학연구원)
  • Published : 2008.12.31

Abstract

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.

Keywords

References

  1. Wagers AJ, Weissman IL: Plasticity of adult stem cells. Cell 2004:116;639-648 https://doi.org/10.1016/S0092-8674(04)00208-9
  2. Mao JJ, Giannobile WV, Helms JA, Hollister SJ, Krebsbach PH, Longaker MT, et al: Craniofacial tissue engineering by stem cells. J Dent Res 2006;85:966-979 https://doi.org/10.1177/154405910608501101
  3. Buranasinsup S, Sila-asna M, Bunyaratvej N, Bunyaratvej A: In vitro osteogenesis from skin-derived precursor cells. Develop Growth Differ 2006;48:263-269 https://doi.org/10.1111/j.1440-169X.2006.00864.x
  4. Shi C, Zhu Y, Su Y, Cheng T: Stem cells and their applications in skin-cell therapy. Trend Biotech 2006;24:48-52 https://doi.org/10.1016/j.tibtech.2005.11.003
  5. Toma JG, Akhavan M, Fernandes KJ, Barnabe-Heider F, Sadikot A, Kaplan DR, et al: Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol 2001:3;778-784 https://doi.org/10.1038/ncb0901-778
  6. Dyce PW, Zhu H, Craig J, Li J: Stem cells with multilineage potential derive from porcine skin. Biochem Biophys Res Commun 2004;316:651-658 https://doi.org/10.1016/j.bbrc.2004.02.093
  7. Kajahn J, Gorjup E, Tiede S, von Briesen H, Paus R, Kruse C, et al: Skin-derived human adult stem cells surprisingly share many features with human pancreatic stem cells. Eur J Cell Biol 2008;87:39-46 https://doi.org/10.1016/j.ejcb.2007.07.004
  8. Toma JG, McKenzie IA, Bagli D, Miller FD: Isolation and characterization of multipotent skin-derived precursors from human skin. Stem Cells 2005;23:727-737 https://doi.org/10.1634/stemcells.2004-0134
  9. Marchesi C, Pluderi M, Colleoni F, Belicchi M, Meregalli M, Farini A, et al: Skin-derived stem cells transplanted into resorbable guides provide functional nerve regeneration after sciatic nerve resection. GLIA 2007;55:425-438 https://doi.org/10.1002/glia.20470
  10. Carlin R, Davis D, Weiss M, Schultz B, Troyer D: Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells. Reprod Biol Endocrinol 2006;4:8-20 https://doi.org/10.1186/1477-7827-4-8
  11. Evans MJ, Kaufman MH: Establishment in culture of pluripotent cells from mouse embryos. Nature 1981;292:154-156 https://doi.org/10.1038/292154a0
  12. Martin GR: Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 1981;78:7634-7638
  13. Scholer HR, Ruppert S, Suzuki N, Chowdhury K, Gruss P: New type of POU domain in germ line-specific protein Oct-4. Nature 1990;344:435-439 https://doi.org/10.1038/344435a0
  14. Avilion AA, Nicolis SK, Pevny LH, Perez L, Vivian N, Lovell- Badge R: Multipotent cell lineages in early mouse development depend on Sox2 function. Genes Dev 2003;17:126-140 https://doi.org/10.1101/gad.224503
  15. Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, et al: Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 2003;113:643-655 https://doi.org/10.1016/S0092-8674(03)00392-1
  16. Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, et al: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 2005;122:947-956 https://doi.org/10.1016/j.cell.2005.08.020
  17. Bucher P, Morel P, Buhler LH: Xenotransplantation: an update on recent progress and future perspectives. Transpl Int 2005;18:894-901 https://doi.org/10.1111/j.1432-2277.2005.00124.x
  18. Schmelzeisen R, Schimming R, Sittinger M: Making bone: implant into tissue-engineered bone for maxillary sinus floor augmentation- a preliminary report. J Cranio Maxillofac Surg 2003;31:34-39 https://doi.org/10.1016/S1010-5182(02)00163-4
  19. Wheeler SL, Holmes RE, Calhoun CJ: Six-year clinical and histologic study of sinus-lift grafts. Int J Oral Maxillofac Implants 1996;11:26-34
  20. Chen F, Feng X, Wu W, Ouyang H, Gao Z, Cheng X, et al: Segmental bone tissue engineering by seeding osteoblast precursor cells into titanium mesh-coral composite scaffolds. Int J Oral Maxillofac Surg 2007;36:822-827 https://doi.org/10.1016/j.ijom.2007.06.019
  21. Yamada Y, Ueda M, Naiki T, Nagasaka T: Tissue-engineered injectable bone regeneration for osseointegrated dental implants. Clin Oral Impl Res 2004;15:589-597 https://doi.org/10.1111/j.1600-0501.2004.01038.x
  22. Fuerst G, Tangl S, Gruber R, Gahleitner A, Sanroman F, Watzek G: Bone formation following sinus grafting with autogenous bone-derived cells and bovine bone mineral in minipigs: preliminary findings. Clin Oral Impl Res 2004;15:733-740 https://doi.org/10.1111/j.1600-0501.2004.01077.x
  23. Springer IN, Nocini PF, Schlegel KA, Santis DD, Park J, Warnke PH, et al: Two techniques for the preparation of cell-scaffold constructs suitable for sinus augmentation: step to clinical application. Tissue Engineering 2006;12:2649-2656 https://doi.org/10.1089/ten.2006.12.2649
  24. Park BW, Hah YS, Kim DY, Kim JR, Byun JH: Osteogenic phenotypes and mineralization of cultured human periosteal-derived cells. Arch Oral Biol 2007;52:983-989 https://doi.org/10.1016/j.archoralbio.2007.04.007
  25. Zou L, Zou X, Chen L, Li H, Mygind T, Kassem M, et al: Multilineage differentiation of porcine bone marrow stromal cells associated with specific gene expression pattern. J Orthop Res 2008;26:56-64 https://doi.org/10.1002/jor.20467