title: [Bone marrow derived cells promoting pre-degeneration of sciatic nerve in vitro].
abstract: OBJECTIVE
  To explore a new method for the pre-degeneration of peripheral nerve in vitro for obtaining many effective Schwann cells so as to provide a large number of seed cells for the research and application of tissue engineered nerves.
  
  
  METHODS
  The bone marrow derived cells (BMDCs) from transgenic green fluorescent protein C57BL/6 mouse and the sciatic nerve segments from the C57BL/6 mouse were co-cultured to prepare the pre-degeneration of sciatic nerve in vitro (experimental group, group A), and only sciatic nerve was cultured (control group, group B). At 7 days after culture, whether BMDCs can permeate into the sciatic nerve in vitro for pre-degeneration was observed by gross and immunohistofluorescence staining. And then Schwann cells were obtained from the sciatic nerves by enzymic digestion and cultured. The cell number was counted, and then the purity of primary Schwann cells was determined using immunohistofluorescence staining and flow cytometer analysis.
  
  
  RESULTS
  At 7 days after pre-degeneration, gross observation showed that enlargement was observed at nerve stumps, and neuroma-like structure formed; the group A was more obvious than group B. Immunohistofluorescence staining showed many BMDCs permeated into the nerve segments, with positive F4/80 staining in group A. After culture, the yield of Schwann cells was (5.59 +/- 0.19) x 10(4) /mg in group A and (3.20 +/-0.21) x 10(4)/mg in group B, showing significant difference (t=2.14, P=0.03). At 48 hours after inoculation, the cells had blue bipolar or tripolar cell nuclei with small size and red soma by immunohistofluorescence staining; fibroblasts were flat polygonal with clear nucleus and nucleolus, showing negative p75NTR staining; and there were few of fibroblasts in group A. The purity of Schwann cells was 88.4% +/- 5.8% in group A and 76.1% +/-3.7% in group B, showing significant difference (t=2.38, P=0.04). And the flow cytometer analysis showed that the purity was 89.6% in group A and 74.9% in group B.
  
  
  CONCLUSIONS
  BMDCs can promote the pre-degeneration of peripheral nerve in vitro, and it is a new method to effectively obtain Schwann cells for tissue engineered nerve.
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Bengbu Medical College, Laboratory of Tissue and Transplant in Anhui Province, Bengbu Anhui, 233003, P.R.China. wumin200207@163.com
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.forename: Xiaopan
authors.initials: X
authors.lastname: Wang
authors.forename: Yuzhou
authors.initials: Y
authors.lastname: Xiao
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D005260:Female; D005434:Flow Cytometry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009416:Nerve Regeneration; D012583:Schwann Cells / Q000166:cytology / Q000502:physiology*; D012584:Sciatic Nerve / Q000166:cytology / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods; D014855:Wallerian Degeneration / Q000473:pathology / Q000503:physiopathology*