Effect of neural stem cell transplantation on neurological function of cerebral hemorrhage rats / 中国组织工程研究

ABSTRACT

BACKGROUND: Exogenous neural stem cells (NSCs) can repair nerve and promote recovery of neurofunction following cerebral hemorrhage.OBJECTIVE: To observe the growth and development of NSCs in vitro, to evaluate the survival, migration and differentiation of transplanted NSCs surrounding hematoma and the possible recovery function of NSCs, and to investigate the repairing effect of NSCs on damaged neurofunction in cerebral hemorrhage model rats.DESIGN: Completely randomized grouping design and controlled animal study,SETTING: Department of Neurosurgery, Huashan Hospital, Fudan University.MATERIALS Eighteen adult healthy male SD rats weighing 280-320 g were provided by Shanghai Animal Center of Chinese Science Academy. BrdU was provided by Neomarkers Company; rat-anti-glial fibrillary acidic protein (GFAP) and rabbit-anti-microtubule-associated protein-2 (MAP-2) by Chemicon Company.METHODS: This study was performed at the Laboratory of Anatomy and Histology & Embryology, Shanghai Medical College,Fudan University from February to December 2006. The NSCs was isolated, cultured, and evaluated from hippocampus of day E14fetal SD rats. Eighteen rats were randomly divided into control group, PBS group and NSC transplantation group. Cerebralhemorrhage rat models were established via injection of autoiogous arterial blood in caudate nucleus. Thirty minutes after model establishment, 5 μ L NSC suspension with the concentration of 2×1011 L-1 was transplanted at four points surrounding hematoma cavity in the NSC transplantation group. Transplantation of PBS and NSCs was the same as autoblood transplantation. Thirty minutes after model establishment, injuries at the four points were performed, and nothing was injected in the control group.MAIN OUTCOME MEASURES: Neurofunction was evaluated with forward limb scale and turning scale just soon after transplantation and at 1, 3, 5, 14, and 28 days after transplantation. Brain was colleted by anesthesia 28 days after model establishment.Differentiation of transplanted NSCs was detected through testing GFAP, MAP-2, and BrdU by using immunohistochemistry.RESULTS: ①Neurofunction scores There was no significant difference 5 days after model establishment (P＞0.05). However, the scores were significantly improved in the NSC transplantation group 14-28 days after model establishment (P＜0.05).②lmmunofluorescent double labeling Apoptosis ceils around hemotoma in the NSC transplantation group were less than those in the PBS group. BrdU and MAP-2 or GFAP-positive ceils were observed in cerebral tissue sections, and this suggested that NSCs could survive, migrate and differentiate in host brain and differentiate into neurons or astrocytes.CONCLUSION: NSC Transplantation contributes to the recovery of neurofunction in cerebral hemorrhage rats through differentiation into neurons or astrocytes.