Abstract
Objective
To examine the survival, migration, integration, differentiation and the expression of various neurotrophic factors of bone-marrow mesenchymal stem cells (BMSCs) transplanted into the vitreous cavity of rats injured by ischemia/reperfusion(I/R).
Methods
The BMSCs were separated from rat marrow using the wall-sticking method, and cultured in vitro to expand. Flow cytometry detected the surface antigens of BMSCs. Ninety-six rats were randomly divided into four groups: normal control injected PBS(C+P),normal control injected BMSCs (C+B), ischemic/reperfusion injected PBS(I/R+P)and ischemic/reperfusion injected BMSCs(I/R+B). After retinal I/R injury was induced in each group by increasing intraocular pressure, 10 µl PBS and BMSC suspensions labeled by red fluorescence CM-Dil were immediately injected into the vitreous cavity. We observed the survival, migration and integration of BMSCs using confocal microscopy. The differentiation and expression of basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) of CM-Dil-labeled BMSCs were detected by immunofluorescent labeling and reserved by confocal microscopy. The expression of mRNA and proteins of bFGF, BDNF and CNTF were assayed by RT-PCR and Western Blot respectively.
Results
After transplantation to normal eyes, BMSCs labeled by CM-Dil were mostly present in the vitreous cavity, and did not migrate. After transplantation to I/R eyes, BMSCs labeled by CM-Dil were mostly present along with the inner limiting membrane. Only a few cells were integrated into the ganglion cell layer. Two or 4 weeks after transplantation, a few BMSCs labeled by CM-Dil were observed to express markers of neuron- neurone specific enolase (NSE), neurofilament (NF) and various neurotrophic factors. The BMSC-injected I/R model eyes showed less reduction in the number of RGCs than that of the I/R eyes with PBS injection.
Conclusions
BMSC transplantation is a valuable neuroprotection tool for the treatment of retina and optic nerve diseases.
Similar content being viewed by others
References
Hood DC, Greenstein V, Frishman L et al (1999) Identifying inner retinal contributions to the human multifocal ERG. Vision Res 39:2285–2291, doi:10.1016/S0042-6989(98)00296-X
Palmowski AM, Sutter EE, Bearse MA Jr, Fung W (1997) Mapping of retinal function in diabetic retinopathy using the multifocal electroretinogram. Invest Ophthalmol Vis Sci 38:2586–2596
Nishida A, Takahashi M, Tanihara H et al (2000) Incorporation and differentiation of hippocampus-derived neural stem cells transplanted in injured adult rat retina. Invest Ophthalmol Vis Sci 41:4268–4274
Takahashi M, Palmer TD, Takahashi J, Gage FH (1998) Widespread integrationand survival of adult-derived neural progenitor cells in the developing optic retina. Mol Cell Neurosci 12:340–348, doi:10.1006/mcne.1998.0721
Kurimoto Y, Shibuki H, Kaneko Y, Ichikawa M, Kurokawa T, Takahashi M, Yoshimura N (2001) Transplantation of adult rat hippocampus-derived neural stem cells into retina injured by transient ischemia. Neurosci Lett 306:57–60, doi:10.1016/S0304-3940(01)01857-2
Guo Y, Saloupis P, Shaw SJ, Rickman DW (2003) Engraftment of adult neural progenitor cells transplanted to rat retina injured by transient Ischemia. Invest Ophthalmol Vis Sci 44:3194–3201, doi:10.1167/iovs.02-0875
Munoz-Elias G, Woodbury D, Black IB (2003) Marrow stromal cells, mitosis, and neuronal differentiation: stem cell and precursor functions. Stem Cells 21:437–448, doi:10.1634/stemcells.21-4-437
Van Hoffelen SJ, Young MJ, Shatos MA, Sakaguchi DS (2003) Incorporation of murine brain progenitor cells into the developing mammalian retina. Invest Ophthalmol Vis Sci 44:426–434, doi:10.1167/iovs.02-0269
Grozdanic SD, Ast AM, Lazic T, Kwon YH, Kardon RH, Sonea IM, Sakaguchi DS (2006) Morphological integration and functional assessment of transplanted neural progenitor cells in healthy and acute ischemic rat eyes. Exp Eye Res 82:597–607, doi:10.1016/j.exer.2005.08.020
Tomita M, Adachi Y, Yamada H, Takahashi K, Kiuchi K, Oyaizu H, Ikebukuro K, Kaneda H, Matsumura M, Ikehara S (2002) Bone marrow-derived stem cells can differentiate into retinal cells in injured rat retina. Stem Cells 20:279–283, doi:10.1634/stemcells.20-4-279
Kicic A, Shen WY, Wilson AS, Constable IJ, Robertson T, Rakoczy PE (2003) Differentiation of marrow stromal cells into photoreceptors in the rat eye. J Neurosci 23:7742–7749
Klassen HJ, Ng TF, Kurimoto Y, Kirov I, Shatos M, Coffey P, Young MJ (2004) Multipotent retinal progenitors express developmental markers, differentiate into retinal neurons, and preserve light-mediated behavior. Invest Ophthalmol Vis Sci 45:4167–4173, doi:10.1167/iovs.04-0511
Chacko DM, Rogers JA, Turner JE, Ahmad I (2000) Survival and differentiation of cultured retinal progenitors transplanted in the subretinal space of the rat. Biochem Biophys Res Commun 268:842–846, doi:10.1006/bbrc.2000.2153
Pressmar S, Ader M, Richard G, Schachner M, Bartsch U (2001) The fate of heterotopically grafted neural precursor cells in the normal and dystrophic adult mouse retina. Invest Ophthalmol Vis Sci 42:3311–3319
Tomita M, Mori T, Maruyama K et al (2006) A comparison of neural differentiation and retinal transplantation with bone marrow-derived cells and retinal progenitor cells. Stem Cells 24:2270–2278, doi:10.1634/stemcells.2005-0507
Dennis JE, Charbord P (2002) Origin and differentiation of human and murine stroma. Stem Cells 20:205–214, doi:10.1634/stemcells.20-3-205
Woodbury D, Emily JS, Darwin JP et al (2000) Adult rat and human bone marrow stromal cell differentiate into neurons. J Neurosci Res 61:364–370, doi:10.1002/1097-4547(20000815)61:4<364::AID-JNR2>3.0.CO;2-C
Sanchez RJ, Song S, Cardozo PF et al (2000) Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 164:247–256, doi:10.1006/exnr.2000.7389
Deng W, Obrocka M, Fischer I et al (2001) In vitro differentiation of human marrow stromal cells into early progenotors of neural cells by conditions that increase intracellular cyclic AMP. Biochem Biophys Res Commun 282:148–152, doi:10.1006/bbrc.2001.4570
Kopen GC, Prockop DJ, Phinney DG (1999) Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci USA 96:10711–10716, doi:10.1073/pnas.96.19.10711
Azizi SA, Stokes D, Augelli BJ, DiGirolamo C, Prockop DJ (1998) Engraftment and migration of human bone marrow stromal cells implanted in the brains of albino rats — Similarities to astrocyte grafts. Proc Natl Acad Sci USA 95:3908–3913, doi:10.1073/pnas.95.7.3908
Yu S, Tanabe T, Dezawa M et al (2006) Effects of bone marrow stromal cell injection in an experimental glaucoma model. Biochem Biophys Res Commun 344:1071–1079, doi:10.1016/j.bbrc.2006.03.231
Sengupta N, Caballero S, Mames RN, Butler JM, Scott EW, Grant MB (2003) The role of adult bone marrow-derived stem cells in choroidal neovascularization. Invest Ophthalmol Vis Sci 44:4908–4913, doi:10.1167/iovs.03-0342
Labouyrie E, Dubus P, Groppi A, Mahon FX, Ferrer J, Parrens M, Reiffers J, de Mascarel A, Merlio JP (1999) Expression of neurotrophins and their receptors in human bone marrow. Am J Pathol 154:405–415
Takai K, Hara J, Matsumoto K, Hosoi G, Osugi Y, Tawa A, Okada S, Nakamura T (1997) Hepatocyte growth factor is constitutively produced by human bone marrow stromal cells and indirectly promotes hematopoiesis. Blood 89:1560–1565
Sensebe L, Deschaseaux M, Li J, Herve P, Charbord P (1997) The broad spectrum of cytokine gene expression by myoid cells from the human marrow microenvironment. Stem Cells 15:133–143
Ye M, Chen S, Wang X, Qi C, Lu G, Liang L, Xu J (2005) Glial cell line-derived neurotrophic factor in bone marrow stromal cells of rat. Neuroreport 16:581–584, doi:10.1097/00001756-200504250-00013
Wislet-Gendebien S, Bruyere F, Hans G, Leprince P, Moonen G, Rogister B (2004) Nestin-positive mesenchymal stem cells favour the astroglial lineage in neural progenitors and stem cells by releasing active BMP4. BMC Neurosci 5:33, doi:10.1186/1471-2202-5-33
Kinnaird T, Stabile E, Burnett MS, Shou M, Lee CW, Barr S, Fuchs S, Epstein SE (2004) Local delivery of marrow-derived stromal cells augments collateral perfusion through paracrine mechanisms. Circulation 109:1543–1549, doi:10.1161/01.CIR.0000124062.31102.57
Dezawa M, Kanno H, Hoshino M, Cho H, Matsumoto N, Itokazu Y, Tajima N, Yamada H, Sawada H, Ishikawa H, Mimura T, Kitada M, Suzuki Y, Ide C (2004) Specific induction of neuronal cells from bone marrow stromal cells and application for autologous transplantation. J Clin Invest 113:1701–1710
Nickells RW (1999) Apoptosis of retinal ganglion cells in glaucoma: an up of the molecular pathways involved in cell death. Surv Ophthalmol 43:151–161, doi:10.1016/S0039-6257(99)00029-6
Kerrigan LA, Zack DJ, Quigley HA, Smith SD, Pease ME (1997) TUNEL-positive ganglion cells in human primary open-angle glaucoma. Arch Ophthalmol 115:1031–1035
Nickells RW, Zack DJ (1996) Apoptosis in ocular disease: a molecular overview. Ophthalmic Genet 17:145–165, doi:10.3109/13816819609057889
Lam TT, Abler AS, Tso M (1999) Apoptosis and caspases after ischemia reperfusion injury in rat retina. Invest Ophthalmol Vis Sci 40:967–975
Honda Y (1996) Celluar and molecular biology of ischemia retina. Nippon Ganka Gakkai Zasshi 100:937–955
Hangai M, Yoshimura N, Yoshida M, Yabuuchi K, Honda Y (1995) Interleukin-1 gene expression in transient retinal ischemia in the rat. Invest Ophthalmol Vis Sci 36:571–578
Hangai M, Yoshimum N, Honda Y (1996) Increased cytokine gene expression in rat retina following transient ischemia. Ophthalmic Res 28:248–254
Cui Q, Lu Q, So KF, Yip HK (1999) CNTF, not other trophic factors, promotes axonal regeneration of axotomized retinal ganglion cells in adult hamsters. Invest Ophthalmol Vis Sci 40:760–766
Sapieha PS, Peltier M, Rendahl KG, Manning WC, Di Polo A (2003) Fibroblast growth factor-2 gene delivery stimulates axon growth by adult retinal ganglion cells after acute optic nerve injury. Mol Cell Neurosci 24:656–672, doi:10.1016/S1044-7431(03)00228-8
Schuettauf F, Vorwerk C, Naskar R, Orlin A, Quinto K, Zurakowski D, Dejneka NS, Klein RL, Meyer EM, Bennett J (2004) Adeno-associated viruses containing bFGF or BDNF are neuroprotective against excitotoxicity. Curr Eye Res 29:379–386, doi:10.1080/02713680490517872
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Na, L., Xiao-rong, L. & Jia-qin, Y. Effects of bone-marrow mesenchymal stem cells transplanted into vitreous cavity of rat injured by ischemia/reperfusion. Graefes Arch Clin Exp Ophthalmol 247, 503–514 (2009). https://doi.org/10.1007/s00417-008-1009-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00417-008-1009-y