[Comparative study on single cell suspension of neural stem cells and neurospheres transplantation for spinal cord injury in rats].
OBJECTIVE
To compare single cell suspension of neural stem cells (NSCs) with neurospheres transplantation for spinal cord injury (SCI) so as to explore the therapeutic effectiveness of two NSCs transplantation methods for SCI.


METHODS
The NSCs were isolated from the spinal cord of adult Sprague Dawley (SD) rats, purified and cultured. At passage 3, the cells were identified by Hoechst33342, Nestin staining, and glial fibrillary acidic protein staining for differentiated cells. Sixty adult SD rats (weighing 230-250 g) were made the SCI models at T10 level with modified Allen method and randomly divided into 3 groups (20 rats in each). The injury sites were treated by injecting 5 microL saline (group A), 5 microL single cells suspensions of NSCs at passage 3 (group B), and 5 microL neurospheres cell suspensions at passage 3 (group C). At preoperation and 3, 7, 14, 21, and 28 days after operation, the locomotor functions of each group were assessed using the Basso, Beattie, and Bresnahan (BBB) rating scale. HE staining was applied to observe the morphology of spinal cord. Subsequently immunofluorescence staining was used to observe microtubule-associated protein 2 (MAP-2).


RESULTS
The cells cultured were NSCs by morphological observation and immunofluorescence staining. After 3 days of modeling surgery, BBB score significantly decreased when compared with preoperative score, and there was no significant difference among 3 groups at 3 and 7 days (P > 0.05). BBB score increased in different degrees with time; at 14, 21, and 28 days, BBB score of groups B and C was better than that of group A, and group C was better than group B, showing significant differences (P < 0.05). HE staining showed that spinal cord structure of group C was more clear than that of groups A and B, and had less scar. There was no significant difference in the number of MAP-2 positive cells among 3 groups at 3 and 7 days (P > 0.05). At 14, 21, and 28 days, the number of MAP-2 positive cells of groups B and C was significantly more than that of group A, and group C was more than group B, showing significant differences (P < 0.05).


CONCLUSIONS
Transplantation of neurospheres suspension compared with single cell can significantly promote NSCs to differentiate into neurons and is conducive to recover the lower extremity function after SCI.
Department of Orthopedics, the First Affiliated Hospital of Anhui Medical University, Hefei Anhui, 230022, PR China.
Lin
L
Zhong
Hui
H
Zhang
Weilu
W
Gao
Zongsheng
Z
Yin
D000818:Animals; D002454:Cell Differentiation; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D008297:Male; D058953:Neural Stem Cells; D009474:Neurons; D051381:Rats; D017207:Rats, Sprague-Dawley; D013119:Spinal Cord Injuries / Q000628:therapy*; D033581:Stem Cell Transplantation
D005904:Glial Fibrillary Acidic Protein