Adipose-Derived Stem Cells Expressing the Neurogenin-2 Promote Functional Recovery After Spinal Cord Injury in Rat
Neurogenin2 (Ngn2) is a proneural gene that directs neuronal differentiation of progenitor cells during development. This study aimed to investigate whether the use of adipose-derived stem cells (ADSCs) over-expressing the Ngn2 transgene (Ngn2–ADSCs) could display the characteristics of neurogenic cells and improve functional recovery in an experimental rat model of SCI. ADSCs from rats were cultured and purified in vitro, followed by genetically modified with the Ngn2 gene. Forty-eight adult female Sprague–Dawley rats were randomly assigned to three groups: the control, ADSCs, and Ngn2–ADSCs groups. The hind-limb motor function of all rats was recorded using the Basso, Beattie, and Bresnahan locomotor rating scale for 8 weeks. Moreover, hematoxylineosin staining and immunohistochemistry were also performed. After neural induction, positive expression rate of NeuN in Ngn2–ADSCs group was upon 90 %. Following transplantation, a great number of ADSCs was found around the center of the injury spinal cord at 1 and 4 weeks, which improved retention of tissue at the lesion site. Ngn2–ADSCs differentiated into neurons, indicated by the expression of neuronal markers, NeuN and Tuj1. Additionally, transplantation of Ngn2–ADSCs upregulated the trophic factors (brain-derived neurotrophic factor and vascular endothelial growth factor), and inhibited the glial scar formation, which was indicated by immunohistochemistry with glial fibrillary acidic protein. Finally, Ngn2–ADSCs-treated animals showed the highest functional recovery among the three groups. These findings suggest that transplantation of Ngn2-overexpressed ADSCs promote the functional recovery from SCI, and improve the local microenvironment of injured cord in a more efficient way than that with ADSCs alone.
KeywordsAdipose-derived stem cells Neurogenin2 Differentiation Spinal cord injury Functional recovery
This study was supported by grants from the National Natural Science Foundation of China (No. 81171147), “Xingwei Project” Key Personal Medical Research Foundation of Health Department of Jiangsu Province (No. RC201156), “Six Categories of Key Person” Research Foundation of Jiangsu Province (No. 069), Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. JX10231801), and the science and technology plan projects of Jintan (JT2014059).
Compliance with Ethical Standards
This study was approved by the Institutional Review Board and Ethics Committee of Nanjing Medical University, and written informed consent was signed by all participants.
Conflict of interest
The authors declare no conflict of interest in this article.
- Arboleda D, Forostyak S, Jendelova P, Marekova D, Amemori T, Pivonkova H, Masinova K, Sykova E (2011) Transplantation of predifferentiated adipose-derived stromal cells for the treatment of spinal cord injury. Cell Mol Neurobiol 31(7):1113–1122. doi: 10.1007/s10571-011-9712-3 CrossRefPubMedGoogle Scholar
- Chen X, Lepier A, Berninger B, Tolkovsky AM, Herbert J (2012) Cultured subventricular zone progenitor cells transduced with neurogenin-2 become mature glutamatergic neurons and integrate into the dentate gyrus. PLoS One 7(2):e31547. doi: 10.1371/journal.pone.0031547 CrossRefPubMedPubMedCentralGoogle Scholar
- Cheng F, Lu XC, Hao HY, Dai XL, Qian TD, Huang BS, Tang LJ, Yu W, Li LX (2014) Neurogenin 2 converts mesenchymal stem cells into a neural precursor fate and improves functional recovery after experimental stroke. Cell Physiol Biochem 33(3):847–858. doi: 10.1159/000358657 CrossRefPubMedGoogle Scholar
- Facchiano F, Fernandez E, Mancarella S, Maira G, Miscusi M, D’Arcangelo D, Cimino-Reale G, Falchetti ML, Capogrossi MC, Pallini R (2002) Promotion of regeneration of corticospinal tract axons in rats with recombinant vascular endothelial growth factor alone and combined with adenovirus coding for this factor. J Neurosurg 97(1):161–168. doi: 10.3171/jns.2002.97.1.0161 CrossRefPubMedGoogle Scholar
- Heinrich C, Blum R, Gascon S, Masserdotti G, Tripathi P, Sanchez R, Tiedt S, Schroeder T, Gotz M, Berninger B (2010) Directing astroglia from the cerebral cortex into subtype specific functional neurons. PLoS Biol 8(5):e1000373. doi: 10.1371/journal.pbio.1000373 CrossRefPubMedPubMedCentralGoogle Scholar
- Hooshmand MJ, Sontag CJ, Uchida N, Tamaki S, Anderson AJ, Cummings BJ (2009) Analysis of host-mediated repair mechanisms after human CNS-stem cell transplantation for spinal cord injury: correlation of engraftment with recovery. PLoS One 4(6):e5871. doi: 10.1371/journal.pone.0005871 CrossRefPubMedPubMedCentralGoogle Scholar
- Iwanami A, Kaneko S, Nakamura M, Kanemura Y, Mori H, Kobayashi S, Yamasaki M, Momoshima S, Ishii H, Ando K, Tanioka Y, Tamaoki N, Nomura T, Toyama Y, Okano H (2005) Transplantation of human neural stem cells for spinal cord injury in primates. J Neurosci Res 80(2):182–190. doi: 10.1002/jnr.20436 CrossRefPubMedGoogle Scholar
- Majumdar MK, Thiede MA, Mosca JD, Moorman M, Gerson SL (1998) Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells. J Cell Physiol 176(1):57–66. doi: 10.1002/(SICI)1097-4652(199807)176:1<57:AID-JCP7>3.0.CO;2-7 CrossRefPubMedGoogle Scholar
- Nori S, Okada Y, Yasuda A, Tsuji O, Takahashi Y, Kobayashi Y, Fujiyoshi K, Koike M, Uchiyama Y, Ikeda E, Toyama Y, Yamanaka S, Nakamura M, Okano H (2011) Grafted human-induced pluripotent stem-cell-derived neurospheres promote motor functional recovery after spinal cord injury in mice. Proc Natl Acad Sci USA 108(40):16825–16830. doi: 10.1073/pnas.1108077108 CrossRefPubMedPubMedCentralGoogle Scholar
- Ohta M, Suzuki Y, Noda T, Ejiri Y, Dezawa M, Kataoka K, Chou H, Ishikawa N, Matsumoto N, Iwashita Y, Mizuta E, Kuno S, Ide C (2004) Bone marrow stromal cells infused into the cerebrospinal fluid promote functional recovery of the injured rat spinal cord with reduced cavity formation. Exp Neurol 187(2):266–278. doi: 10.1016/j.expneurol.2004.01.021 CrossRefPubMedGoogle Scholar
- Portron S, Merceron C, Gauthier O, Lesoeur J, Sourice S, Masson M, Fellah BH, Geffroy O, Lallemand E, Weiss P, Guicheux J, Vinatier C (2013) Effects of in vitro low oxygen tension preconditioning of adipose stromal cells on their in vivo chondrogenic potential: application in cartilage tissue repair. PLoS One 8(4):e62368. doi: 10.1371/journal.pone.0062368 CrossRefPubMedPubMedCentralGoogle Scholar
- Schreiber J, Schachner M, Schumacher U, Lorke DE (2013) Extracellular matrix alterations, accelerated leukocyte infiltration and enhanced axonal sprouting after spinal cord hemisection in tenascin-C-deficient mice. Acta Histochem 115(8):865–878. doi: 10.1016/j.acthis.2013.04.009 CrossRefPubMedGoogle Scholar
- Thoma EC, Wischmeyer E, Offen N, Maurus K, Siren AL, Schartl M, Wagner TU (2012) Ectopic expression of neurogenin 2 alone is sufficient to induce differentiation of embryonic stem cells into mature neurons. PLoS One 7(6):e38651. doi: 10.1371/journal.pone.0038651 CrossRefPubMedPubMedCentralGoogle Scholar