[Experimental study on transplantation of embryonic stem cells in treating spinal cord injury].
OBJECTIVE
To observe the effect of transplantation of embryonic stem cell (ES) on neurological functional recovery of injured spinal cord in adult mouse.


METHODS
The ES cells were cultured and induced in vitro. Fifty C57/BL6J mice were made animal model of semi-cut mice of T9,10. The ES cell-derived neural precursors cells were transplanted into the vertebral canal around injured spinal cord semi-cut mice. Twenty-eight C57/BL6J mice were randomly divided into three groups: sham operation group(group A, n=9), operation/cell group (group B, n = 10), and operation/DMEM group (group C, n = 9). RT-PCR analysis, X-gal staining and immunofluorescence were used to observe the cells survival and differentiation in the spinal crod. BBB test was performed to study functional improvement.


RESULTS
ES cells induced and cultured in vitro displayed clonal growth with circle or ovoid shape and had one or more nucleoli. RT-PCR result showed that the induced ES cells expressed mRNA of Nestin and microtubule-associated protein, but did not express glial fibrillory acidic protein (GFAP). There was statistically significant difference in BBB scoring between group A and groups B, C after operation (P <0.01). There was statistically significant difference in BBB scoring at 1, 2 and 4 weeks of operation (P < 0.01), but no statistically significant difference at 6 and 8 weeks of operation between groups B and C (P>0.05). The X-gal staining results were positive in group B and negative in groups A and C. The immunoflurescence result showed neurofilament green fluor and no expression of GFAP in injured spinal cord region.


CONCLUSIONS
After transplantation, ES cell-derived cells can survive, transfer into the injury position, and differentiate into neurons, but spinal cord function has no obvious improvement.
Yang
Jianhua
J
Orthopedics Department, the First Affiliated Hospital, Jiamusi University, Jiamusi Heilongjiang, 154002, P. R. China. jianhua01@163.com
Li
Changde
C
Zhai
Raosheng
R
D000818:Animals; D002454:Cell Differentiation; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D006614:Hindlimb / Q000503:physiopathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011897:Random Allocation; D020127:Recovery of Function; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000473:pathology / Q000503:physiopathology / Q000601:surgery*; D033581:Stem Cell Transplantation
D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins