title: Differentiation of human bone marrow precursor cells into neuronal-like cells after transplantation into canine spinal cord organotypic slice cultures.
abstract: BACKGROUND
  Treatments to regenerate different tissue involving the transplantation of bone marrow derived mesenchymal precursor cells are anticipated. Using an alternative methods, in vitro organotypic slice culture method, would be useful to transplant cells and assessing the effects. This study was to determine the possibility of differentiating human bone marrow precursor cells into cells of the neuronal lineage by transplanting into canine spinal cord organotypic slice cultures.
  
  
  METHODS
  Bone marrow aspirates were obtained from posterior superior iliac spine (PSIS) of patients that had undergone spinal fusion due to a degenerative spinal disorder. For cell imaging, mesenchymal precursor cells (MPCs) were pre-stained with PKH-26 just before transplantation to canine spinal cord slices. Canine spinal cord tissues were obtained from three adult beagle dogs. Spinal cords were cut into transverse slices of 1 mm using tissue chopper. Two slices were transferred into 6-well plate containing 3 ml DMEM with antibiotics. Prepared MPCs (1×10(4)) were transplanted into spinal cord slices. On days 0, 3, 7, 14, MPCs were observed for morphological changes and expression of neuronal markers through immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  The morphological study showed: spherical cells in the control and experiment groups on day 0; and on day 3, cells in the control group had one or two thick, short processes and ones in the experiment group had three or four thin, long processes. On day 7, these variously-sized processes contacted each other in the experiment group, but showed typical spindle-shaped cells in the control group. Immunofluorescence showed that PKH-26(+) MPCs stained positive for NeuN(+) and GFAP(+) in experimental group only. Also RT-PCR showed weak expression of β-tubulin III and GFAP.
  
  
  CONCLUSIONS
  Human bone marrow mesenchymal precursor cells (hMPCs) have the potential to differentiate into the neuronal like cells in this canine spinal cord organotypic slice culture model. Furthermore, these findings suggested the possibility that these cells can be utilized to treat patients with spinal cord injuries.
authors.affiliation: Department of Orthopaedics, Shenzhen Second People's Hospital, Shenzhen, Guangdong, China.
authors.forename: Zhi-qiang
authors.initials: ZQ
authors.lastname: Fei
authors.forename: Jian-yi
authors.initials: JY
authors.lastname: Xiong
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Hui-yong
authors.initials: HY
authors.lastname: Shen
authors.forename: Ngo
authors.initials: N
authors.lastname: Stephanie
authors.forename: Wang
authors.initials: W
authors.lastname: Jeffrey
authors.forename: Da-ping
authors.initials: DP
authors.lastname: Wang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004285:Dogs; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D013116:Spinal Cord / Q000166:cytology*