title: Differentiation of nerve-derived adult pluripotent stem cells into osteoblastic and endothelial cells.
abstract: Stem cell-involved tissue engineering has gained dramatic attention as a therapeutic strategy for tissue regeneration including bone repair. However, the currently available possibilities to use embryonic stem cells and induced pluripotent stem cells (iPCs) face potential ethical issues, as well as risks of malignant transformation and immune rejection. Recently identified peripheral nerve-derived adult pluripotent stem cells (NEDAPS) that quickly proliferate after exposure to bone morphogenetic protein-2 (BMP-2) or nerve trauma and exhibit many embryonic stem cell characteristics may provide an attractive source cells for a variety of regenerative therapies.
  
  
  
  The study aimed to examine the differentiation potential of the NEDAPS cells into osteoblastic cells and endothelial cells.
  
  
  
  An in vitro investigation was undertaken to induce mouse NEDAPS cells into the phenotypes of osteoblastic and endothelial cells.
  
  
  
  NEDAPS cells were isolated from low-dose BMP-2-exposed mouse sciatic nerves by collagenase and trypsin extraction. The cells were cultured in a stem cell maintenance medium, and the expression of KLF4, Sox2, c-Myc, and Oct4 before differentiation was confirmed. The cells were then subcultured in a complete osteogenic cell induction medium or endothelial cell growth medium, respectively, at 37°C and 5%CO2atmosphere. Histologic, morphologic, and molecular assessments were performed 7 days later.
  
  
  
  The cells propagated in complete osteogenic medium for 7 days showed strong staining for type I collagen and alkaline phosphatase, suggesting the structural and functional properties of the osteoblastic cells. Further, real-time polymerase chain reaction (RT-PCR) revealed a significant expression of the osteoblast markers osteocalcin, osteopontin, and type I collagen. Similarly, the cells in endothelial growth medium were successfully differentiated into cobblestone-shaped endothelial cells expressing vascular endothelial growth factor (VEGF) receptors Flk-1 and Flt-1 demonstrated by RT-PCR.
  
  
  
  NEDAPS cells are readily induced to osteoblastic and endothelial cells, suggesting therapeutic potential for bone repair and other regenerative therapies.
authors.affiliation: Department of Orthopaedic Surgery, University of Kansas School of Medicine-Wichita, 929 N St Francis St, Wichita, KS 67214, USA; Department of Biological Sciences, Wichita State University, 1845 Fairmount St, Wichita, KS 67260, USA. Electronic address: shang-you.yang@wichita.edu.
authors.forename: Shang-You
authors.initials: SY
authors.lastname: Yang
authors.affiliation: Department of Orthopaedic Surgery, University of Kansas School of Medicine-Wichita, 929 N St Francis St, Wichita, KS 67214, USA.
authors.forename: Nora
authors.initials: N
authors.lastname: Strong
authors.affiliation: Department of Biological Sciences, Wichita State University, 1845 Fairmount St, Wichita, KS 67260, USA.
authors.forename: Xuan
authors.initials: X
authors.lastname: Gong
authors.affiliation: Department of Orthopaedic Surgery, University of Kansas School of Medicine-Wichita, 929 N St Francis St, Wichita, KS 67214, USA. Electronic address: heggeness@kumc.edu.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Heggeness
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008807:Mice, Inbred BALB C; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D024042:Collagen Type I; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D042461:Vascular Endothelial Growth Factor A; D015675:Osteocalcin; D053495:Osteopontin; C501163:Flt1 protein, mouse; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: BMP-2; Cell differentiation; Endothelial cells; Mouse; Nerve-derived adult stem cells; Osteoblast; Osteonectin; Osteopontin; Peripheral nerve; Tissue engineering