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Umbilical Cord Blood Stem Cell Mediated Downregulation of Fas Improves Functional Recovery of Rats after Spinal Cord Injury

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Abstract

Human umbilical cord blood stem cells (hUCB), due to their primitive nature and ability to develop into nonhematopoietic cells of various tissue lineages, represent a potentially useful source for cell-based therapies after spinal cord injury (SCI). To evaluate their therapeutic potential, hUCB were stereotactically transplanted into the injury epicenter, one week after SCI in rats. Our results show the presence of a substantial number of surviving hUCB in the injured spinal cord up to five weeks after transplantation. Three weeks after SCI, apoptotic cells were found especially in the dorsal white matter and gray matter, which are positive for both neuron and oligodendrocyte markers. Expression of Fas on both neurons and oligodendrocytes was efficiently downregulated by hUCB. This ultimately resulted in downregulation of caspase-3 extrinsic pathway proteins involving increased expression of FLIP, XIAP and inhibition of PARP cleavage. In hUCB-treated rats, the PI3K/Akt pathway was also involved in antiapoptotic actions. Further, structural integrity of the cytoskeletal proteins α-tubulin, MAP2A&2B and NF-200 has been preserved in hUCB treatments. The behavioral scores of hind limbs of hUCB-treated rats improved significantly than those of the injured group, showing functional recovery. Taken together, our results indicate that hUCB-mediated downregulation of Fas and caspases leads to functional recovery of hind limbs of rats after SCI.

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Abbreviations

APC:

Adenomatous polyposis coli

BBB:

Basso Beattie Bresnahan locomotor scoring

BCA:

Bicinchoninic acid

BDNF:

Brain derived neurotrophic factor

bFGF:

Basic fibroblast growth factor

CHAPS:

3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate

CNPase:

2′,3′-Cyclicnucleotide-3′-phosphodiesterase

DAB:

Diaminobenzidine

DAPI:

4′,6-Diamidino-2-phenylindole dihydrochloride

DPI:

Days post injury

DTT:

Dithiothreitol

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

FLIP:

FLICE-inhibitory protein

hEGF:

Human epidermal growth factor

HRP:

Horseradish peroxidase

hUCB:

Human umbilical cord blood stem cells

MAP2A&2B:

Microtubule associated protein 2A & 2B

MBP:

Myelin Basic Protein

β-NGF:

Beta-nerve growth factor

NF-200:

Neurofilament H-200 kDa

NT-3:

Neurotrophic hormone-3

PARP:

Poly [ADP-ribose] polymerase

PBS:

Phosphate buffered saline

PMSF:

Phenyl methane sulfonyl fluoride

RA:

Retinoic acid

SCI:

Spinal cord injury

XIAP:

X-linked inhibitor of apoptosis protein

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Acknowledgements

This research was supported by National Cancer Institute Grant CA 75557, CA 92393, CA 95058, CA 116708, N.I.N.D.S. NS47699 and NS57529, and Caterpillar, Inc., OSF Saint Francis, Inc., Peoria, IL (to J.S.R.). We thank Noorjehan Ali for technical assistance. We thank Shellee Abraham for manuscript preparation and Diana Meister and Sushma Jasti for manuscript review.

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Authors and Affiliations

  1. Department of Cancer Biology and Pharmacology, University of Illinois College of Medicine, Peoria, IL, 61656, USA

    Venkata Ramesh Dasari, Liang Li & Jasti S. Rao

  2. Department of Neurosurgery, University of Illinois College of Medicine at Peoria, One Illini Drive, Peoria, IL, 61605, USA

    Daniel G. Spomar, Jasti S. Rao & Dzung H. Dinh

  3. Department of Pathology, University of Illinois College of Medicine at Peoria, Peoria, IL, 61656, USA

    Meena Gujrati

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  1. Venkata Ramesh Dasari
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Correspondence to Dzung H. Dinh.

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Dasari, V.R., Spomar, D.G., Li, L. et al. Umbilical Cord Blood Stem Cell Mediated Downregulation of Fas Improves Functional Recovery of Rats after Spinal Cord Injury. Neurochem Res 33, 134–149 (2008). https://doi.org/10.1007/s11064-007-9426-6

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  • DOI: https://doi.org/10.1007/s11064-007-9426-6

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