Functional recovery after rhesus monkey spinal cord injury by transplantation of bone marrow mesenchymal-stem cell-derived neurons

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Author:
DENG Yu-bin()
YUAN Qing-tao()
LIU Xiao-gang()
LIU Xiao-lin()
LIU Yu()
LIU Zu-guo()
ZHANG Cheng()
Journal Title:
CHINESE MEDICAL JOURNAL
Issue:
Volume 118, Issue 18, 2005
DOI:
Key Word:
mesenchymal stem cell;neuron;spinal cord injury;cell transplantation;rhesus

Abstract´╝Ü Background The treatment of spinal cord injury is still a challenge. This study aimed at evaluating the therapeutical effectiveness of neurons derived form mesenchymal stem cells (MSCs) for spinal cord injury.Methods In this study, rhesus MSCs were isolated and induced by cryptotanshinone in vitro and then a process of RT-PCR was used to detect the expression of glutamic acid decarboxylase (GAD) gene. The induced MSCs were tagged with Hoechst 33342 and injected into the injury site of rhesus spinal cord made by the modified Allen method. Following that, behavior analysis was made after 1 week, 1 month, 2 months and 3 months. After 3 months, true blue chloride retrograde tracing study was also used to evaluate the re-establishment of axons pathway and the hematoxylin-eosin (HE) staining and immunohistochemistry were performed after the animals had been killed.Results In this study, the expression of mRNA of GAD gene could be found in the induced MSCs but not in primitive MSCs and immunohistochemistry could also confirm that rhesus MSCs could be induced and differentiated into neurons. Behavior analysis showed that the experimental animals restored the function of spinal cord up to grade 2-3 of Tarlov classification. Retrograde tracing study showed that true blue chollide could be found in the rostral thoracic spinal cords, red nucleus and sensory-motor cortex.Conclusions These results suggest that the transplantation is safe and effective.

  • [1]Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cell.Science 1999;284:143-147.
  • [2]Gerson SL.Mesenchymal stem cells: no longer second class marrow citizens.Nat Med 1999;5:262-264.
  • [3]Jaiswal N, Haynesworth SE, Caplan AI, et al. Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro.J Cell Biochem 1997;64:295-312.
  • [4]Wakitani S, Saito T, Caplan AI.Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacutidine.Muscle Nerve 1995;18:1417-1426.
  • [5]Schwartz RE. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells.J Clin Invest 2002;109:1291-1302.
  • [6]Kopen GC, Prockop DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains.Proc Natl Acad Sci U S A 1999;96:10711-10716.
  • [7]Woodbury D, Schwaiz EJ, Prockop DJ, et al.Adult rat and human bone marrow stromal cells differentiate into neurons.J Neurosci Res 2000;61:364-370.
  • [8]Sanchez-Ramos J, Song S, Cardozo-Pelaez F, et al.Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 2000;164:247-256.
  • [9]Deng W, Obrocka M, Fischer I, et al. In vitro differentiation of human marrow stromal cells into early progenitors of neural cells by conditions that increase intracellular cyclic AMP.Biochem Biophys Res Commun 2001;282:148-152.
  • [10]Conget PA, Minguell JJ. Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol 1999;181:67-73.
  • [11]Xiao QZ. Adult rat and human bone marrow mesenchymal stem cells differentiate into neurons with Musk's polypeptide. Chin J Pathophysiol (chin) 2002;18:1179-1182.
  • [12]Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Haematol 1976;4:267-274.
  • [13]Mitsushima D. Role of glutamic acid decarboxylase in the prepubertal inhibition of the luteinizing hormone releasing hormone release in female rhesus monkeys. J Neurosci 1996;16:2563-2673.
  • [14]Naslund TC, Hollier LH, Money SR, et al. Protecting the ischemic spinal cord during aortic clamping. Ann Surg 1992:215:409-415.
  • [15]Granke K, Hollier LH, Zdrahal P, et al. Paraplegia following thoracic aorta cross-clamping in the canine model: effect of interventional therapy . J Vasc Surg 1991;13:615-621.
  • [16]Wisselink W, Money SR, Crockett DE, et al. Ischemia reperfusion injury of the spinal cord: protective effect of the hydroxyl radical scavenger dimethylthiourea. J Vasc Surg 1994;20:444-450.
  • [17]Gledhill RF, Harrison BM, MacDonald WI. Pattern of remyelination in the CNS. Nature 1973; 244:443-444.
  • [18]Griffiths IR, Mcculloch MC. Nerve fibres in spinal cord impact injuries. Part 1. Changes in the myelin sheath during the initial 5 weeks. J Neurol Sci 1983;58:335-349.
  • [19]Blight AR. Delayed demyelination and macrophage invasion: a candidate for secondary cell damage in spinal cord injury. Cent Nerv Syst Trauma 1985;2:299-315.
  • [20]Bunge RP, Puckett WR, Hiester ED. Observations on the pathology of several types of human spinal cord injury, with emphasis on the astrocyte response to penetrating injuries. Adv Neurol 1997;72:305-315.
  • [21]van den Bos C, Mosca JD, Winkles J, et al. Human mesenchymal stem cells respond to fibroblast growth factors. Hum Cell 1997;10:45-50.
  • [22]Mezey E, Chandross KJ, Harta G, et al. Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow. Science 2000; 290:1779-1782.
  • [23]Brazelton TR, Rossi FM, Keshet GI, et al. From marrow to brain: expression of neuronal phenotypes in adult mice.Science 2000;290:1775-1779.
  • [24]Chopp M, Zheng XH, Li Y, et al. Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation. Neuroreport 2000;11:3001-3005.
  • [25]Lu D, Mahmood A, Wang L, et al. Adult bone marrow stromal cells administered intravenously to rats after traumatic brain injury migrate into brain and improve neurological outcome. Neuroreport 2001;12:559-563.
  • [26]Ramer MS, Priestley JV, McMahon SB. Functional regeneration of sensory axons into the adult spinal cord. Nature 2000;403:312-316.
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