Human induced pluripotent stem cell-derived neural stem cells survive, migrate, differentiate, and improve neurologic function in a rat model of middle cerebral artery occlusion

被引:110
作者
Yuan, Ting [1 ]
Liao, Wei [3 ]
Feng, Nian-Hua [4 ]
Lou, Yuan-Lei [4 ]
Niu, Xin [1 ]
Zhang, Ai-Jun [2 ]
Wang, Yang [1 ,4 ]
Deng, Zhi-Feng [2 ,3 ]
机构
[1] Shanghai Jiao Tong Univ, Affiliated Peoples Hosp 6, Dept Orthopaed Surg, Shanghai 200233, Peoples R China
[2] Shanghai Jiao Tong Univ, Affiliated Peoples Hosp 6, Dept Neurosurg, Shanghai 200233, Peoples R China
[3] Nanchang Univ, Affiliated Hosp 2, Dept Neurosurg, Nanchang 330006, Peoples R China
[4] Nanchang Univ, Affiliated Hosp 1, Inst Urol, Nanchang 330006, Peoples R China
关键词
Induced pluripotent stem cell; Stoke; Neural stem cell; Middle cerebral artery occlusion; IN-VITRO DIFFERENTIATION; CORD BLOOD-CELLS; PRECURSOR CELLS; RETINOIC-ACID; ENRICHED ENVIRONMENT; STROKE; NEURONS; BRAIN; TRANSPLANTATION; ANGIOGENESIS;
D O I
10.1186/scrt224
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Introduction: Stroke is a major cause of permanent neurologic damage, with few effective treatments available to restore lost function. Induced pluripotent stem cells (iPSCs) have the potential to generate all cell types in vitro and can be generated from a stroke patient. Therefore, iPSCs are attractive donor sources of genetically identical "patient-specific" cells to hold promise in therapy for stroke. In the present study, we established a four-stage culture system by using serum-free medium and retinoic acid (RA) to differentiate iPSCs into neural stem cells (NSCs) effectively and stably. Our hypothesis was that iPSC-derived NSCs would survive, migrate, and differentiate in vivo, and improve neurologic function after transplantation into the brains of rats with ischemic stroke. Methods: Human iPSCs (iPS-S-01) and human ESCs (HuES17) were used to differentiate into NSCs by using our four-stage culture system. iPSCs and differentiated NSCs were characterized by immunocytochemistry staining and reverse transcription-polymerase chain reaction (RT-PCR) analysis. After establishment of focal cerebral ischemia with occlusion of the middle cerebral artery (MCA) and cell transplantation, animals were killed at 1 week and 2 weeks to analyze survival, migration, and differentiation of implanted cells in brain tissue. Animal behavior was evaluated via rope grabbing, beam walking, and Morris water maze tests. Results: iPSCs were efficiently induced into NSCs by using a newly established four-stage induction system in vitro. iPSCs expressed pluripotency-associated genes Oct4, Sox2, and Nanog before NSC differentiation. The iPSC-derived NSCs spontaneously differentiated into neurons and astrocytes, which highly express beta-tubulin and glial fibrillary acidic protein (GFAP), respectively. On transplantation into the striatum, CM-DiI labeled iPSC-derived NSCs were found to migrate into the ischemia area at 1 week and 2 weeks, and animal-function recovery was significantly improved in comparison with control groups at 3 weeks. Conclusions: The four-stage induction system is stable and effective to culture, differentiate, and induce iPSCs to NSCs by using serum-free medium combined with retinoic acid (RA). Implanted iPSC-derived NSCs were able to survive, migrate into the ischemic brain area to differentiate into mature neural cells, and seem to have potential to restore lost neurologic function from damage due to stroke in a rat model.
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页数:10
相关论文
共 47 条
[1]   Neural differentiation from human embryonic stem cells in a defined adherent culture condition [J].
Baharvand, Hossein ;
Mehrjardi, Narges-Zare ;
Hatami, Maryam ;
Kiani, Sahar ;
Rao, Mahendra ;
Haghighi, Mahdi-Montazer .
INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY, 2007, 51 (05) :371-378
[2]   EMBRYONIC STEM-CELLS EXPRESS NEURONAL PROPERTIES IN-VITRO [J].
BAIN, G ;
KITCHENS, D ;
YAO, M ;
HUETTNER, JE ;
GOTTLIEB, DI .
DEVELOPMENTAL BIOLOGY, 1995, 168 (02) :342-357
[3]   The potential benefit of stem cell therapy after stroke: an update [J].
Banerjee, Soma ;
Williamson, Deborah A. ;
Habib, Nagy ;
Chataway, Jeremy .
VASCULAR HEALTH AND RISK MANAGEMENT, 2012, 8 :569-580
[4]   Stem cell therapy: A clinical trial of stroke [J].
Bhasin, Ashu ;
Srivastava, M. V. Padma ;
Mohanty, Sujata ;
Bhatia, Rohit ;
Kumaran, Senthil S. ;
Bose, Sushmita .
CLINICAL NEUROLOGY AND NEUROSURGERY, 2013, 115 (07) :1003-1008
[5]   Embryonic stem cells develop into functional dopaminergic neurons after transplantation in a Parkinson rat model [J].
Björklund, LM ;
Sánchez-Pernaute, R ;
Chung, SM ;
Andersson, T ;
Chen, IYC ;
McNaught, KS ;
Brownell, AL ;
Jenkins, BG ;
Wahlestedt, C ;
Kim, KS ;
Isacson, O .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (04) :2344-2349
[6]  
Bonnamain V, 2012, FRONT CELL NEUROSCI, V6
[7]   Intravenous administration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats [J].
Chen, JL ;
Zhang, ZG ;
Li, Y ;
Wang, L ;
Xu, YX ;
Gautam, SC ;
Lu, M ;
Zhu, Z ;
Chopp, M .
CIRCULATION RESEARCH, 2003, 92 (06) :692-699
[8]   Functional Improvement of Focal Cerebral Ischemia Injury by Subdural Transplantation of Induced Pluripotent Stem Cells with Fibrin Glue [J].
Chen, Shih-Jen ;
Chang, Chia-Ming ;
Tsai, Shen-Kou ;
Chang, Yuh-Lih ;
Chou, Shih-Jie ;
Huang, Shiang-Suo ;
Tai, Lung-Kuo ;
Chen, Yu-Chih ;
Ku, Hung-Hai ;
Li, Hsin-Yang ;
Chiou, Shih-Hwa .
STEM CELLS AND DEVELOPMENT, 2010, 19 (11) :1757-1767
[9]   Human neural stem cells can migrate, differentiate, and integrate after intravenous transplantation in adult rats with transient forebrain ischemia [J].
Chu, K ;
Kim, M ;
Jeong, SW ;
Kim, SU ;
Yoon, BW .
NEUROSCIENCE LETTERS, 2003, 343 (02) :129-133
[10]   COMBINATION TREATMENT OF STROKE WITH SUB-THERAPEUTIC DOSES OF SIMVASTATIN AND HUMAN UMBILICAL CORD BLOOD CELLS ENHANCES VASCULAR REMODELING AND IMPROVES FUNCTIONAL OUTCOME [J].
Cui, X. ;
Chopp, M. ;
Zacharek, A. ;
Dai, J. ;
Zhang, C. ;
Yan, T. ;
Ning, R. ;
Roberts, C. ;
Shehadah, A. ;
Kuzmin-Nichols, N. ;
Sanberg, C. D. ;
Chen, J. .
NEUROSCIENCE, 2012, 227 :223-231