Predifferentiated embryonic stem cells promote functional recovery after spinal cord compressive injury.
We tested the effects of mouse embryonic stem cells (mES) grafts in mice spinal cord injury (SCI). Young adult female C57/Bl6 mice were subjected to laminectomy at T9 and 1-minute compression of the spinal cord with a vascular clip. Four groups were analyzed: laminectomy (Sham), injured (SCI), vehicle (DMEM), and mES-treated (EST). mES pre-differentiated with retinoic acid were injected (8 x 10(5) cells/2 microl) into the lesion epicenter, 10 min after SCI. Basso mouse scale (BMS) and Global mobility test (GMT) were assessed weekly up to 8 weeks, when morphological analyses were performed. GMT analysis showed that EST animals moved faster (10.73+/-0.9076, +/-SEM) than SCI (5.581+/-0.2905) and DMEM (5.705+/-0.2848), but slower than Sham animals (15.80+/-0.3887, p<0.001). By BMS, EST animals reached the final phase of locomotor recovery (3.872+/-0.7112, p<0.01), while animals of the SCI and DMEM groups improved to an intermediate phase (2.037+/-0.3994 and 2.111+/-0.3889, respectively). White matter area and number of myelinated nerve fibers were greater in EST (46.80+/-1.24 and 279.4+/-16.33, respectively) than the SCI group (39.97+/-0.925 and 81.39+/-8.078, p<0.05, respectively). EST group also presented better G-ratio values when compared with SCI group (p<0.001). Immunohistochemical revealed the differentiation of transplanted cells into astrocytes, oligodendrocytes, and Schwann cells, indicating an integration of transplanted cells with host tissue. Ultrastructural analysis showed, in the EST group, better tissue preservation and more remyelination by oligodendrocytes and Schwann cells than the other groups. Our results indicate that acute transplantation of predifferentiated mES into the injured spinal cord increased the spared white matter and number of nerve fibers, improving locomotor function.
Marques
Suelen Adriani
SA
Laboratório de Regeneração Neural e Função, Instituto de Biologia, Universidade Federal Fluminense, Brazil. suelen_marques@yahoo.com
Almeida
Fernanda Martins
FM
Fernandes
Aline Marie
AM
dos Santos Souza
Cleide
C
Cadilhe
Daniel Veloso
DV
Rehen
Stevens Kastrup
SK
Martinez
Ana Maria Blanco
AM
D000704:Analysis of Variance; D000818:Animals; D001522:Behavior, Animal; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000502:physiology*; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009043:Motor Activity / Q000502:physiology; D004676:Myelin Basic Protein / Q000378:metabolism; D009413:Nerve Fibers, Myelinated / Q000473:pathology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009457:Neuroglia / Q000502:physiology / Q000648:ultrastructure; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D020127:Recovery of Function / Q000502:physiology*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D012583:Schwann Cells / Q000502:physiology / Q000648:ultrastructure; D013117:Spinal Cord Compression / Q000473:pathology / Q000503:physiopathology* / Q000601:surgery*; D033581:Stem Cell Transplantation
C489787:MAP2 protein, rat; D008869:Microtubule-Associated Proteins; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors