title: Organotypic human spinal cord slice culture as an alternative to direct transplantation of human bone marrow precursor cells for treating spinal cord injury.
abstract: OBJECTIVE
  To determine the possibility of differentiation of human bone marrow-derived mesenchymal precursor cells (BMDMPCs) into neuronal lineage cells using a human spinal cord organotypic slice coculture technique as an alternative to an in vivo human study.
  
  
  METHODS
  Human BMDMPCs were stained with PKH-26 dye before transplantation into 12 human spinal cord slices. In the control group, BMDMPCs were embedded into one spinal cord-free six-well plate containing media. The morphologic differentiation of the transplanted BMDMPCs were observed at 0, 3, 7, and 14 days. Neuroglial differentiation was identified with immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  Spherical cells were seen in both groups at day 0. On days 7 and 14, cells developed one or two thick, short processes and typical spindle-shaped cells in the control group and three to five thin, long processes and neuron-like cells in the experimental group. Immunohistochemistry showed double-stained cells with PKH-26 dye (positive) and vimentin (positive), PKH-26 (positive) and neuronal nuclei (NeuN) (positive), and PKH-26 (positive) and glial fibrillary acidic protein (GFAP) (positive) in the experimental group only. RT-PCR showed weak expression of tyrosine kinase A, NeuN, β-tubulin III, and GFAP in the experimental group.
  
  
  CONCLUSIONS
  Organotypic human spinal cord slice culture may be a useful method to verify the neuroglial differentiation of human BMDMPCs as an alternative to a direct human study.
authors: Department of Animal Source, Jeju National University, College of Life Sciences, Jeju, Korea.
authors: Dong-Kee
authors: DK
authors: Jeong
authors: Cyrus E
authors: CE
authors: Taghavi
authors: Kyung-Jin
authors: KJ
authors: Song
authors: Kwang-Bok
authors: KB
authors: Lee
authors: Hyun-Wook
authors: HW
authors: Kang