Hostname: page-component-76fb5796d-x4r87 Total loading time: 0 Render date: 2024-04-25T17:25:57.229Z Has data issue: false hasContentIssue false
  • English
  • Français

Functional and morphological analysis of the subretinal injection of retinal pigment epithelium cells

Published online by Cambridge University Press:  01 March 2012

MAREN ENGELHARDT ,
CHINATSU TOSHA ,
VANDA S. LOPES ,
BRYAN CHEN ,
LISA NGUYEN ,
STEVEN NUSINOWITZ  and
DAVID S. WILLIAMS
MAREN ENGELHARDT
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California Department of Anatomy, Institute of Neuroanatomy, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany
CHINATSU TOSHA
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California
VANDA S. LOPES
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California Centre of Ophthalmology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal
BRYAN CHEN
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California
LISA NGUYEN
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California
STEVEN NUSINOWITZ*
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California
DAVID S. WILLIAMS*
Affiliation:
Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, California Department of Neurobiology, David Geffen School of Medicine, University of California, Los Angeles, California
*
‡Address correspondence and reprint requests to: Dr. David S. Williams or Dr. Steven Nusinowitz, Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095. E-mail: dswilliams@ucla.edu or nusinowitz@jsei.ucla.edu
‡Address correspondence and reprint requests to: Dr. David S. Williams or Dr. Steven Nusinowitz, Department of Ophthalmology, Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095. E-mail: dswilliams@ucla.edu or nusinowitz@jsei.ucla.edu
Get access Rights & Permissions [Opens in a new window]

Abstract

Replacement of retinal pigment epithelium (RPE) cells by transplantation is a potential treatment for some retinal degenerations. Here, we used a combination of invasive and noninvasive methods to characterize the structural and functional consequences of subretinal injection of RPE cells. Pigmented cells from primary cultures were injected into albino mice. Recovery was monitored over 8 weeks by fundus imaging, spectral domain optical coherence tomography (sdOCT), histology, and electroretinography (ERG). sdOCT showed that retinal reattachment was nearly complete by 1 week. ERG response amplitudes were reduced after injection, with cone-mediated function then recovering better than rod function. Photoreceptor cell loss was evident by sdOCT and histology, near the site of injection, and is likely to have been the main cause of incomplete recovery. With microscopy, injected cells were identified by the presence of apical melanosomes. They either established contact with Bruch’s membrane, and thus became part of the RPE monolayer, or were located on the apical surface of the host’s cells, resulting in apposition of the basal surface of the injected cell with the apical surface of the host cell and the formation of a series of desmosomal junctions. RPE cell density was not increased, indicating that the incorporation of an injected cell into the RPE monolayer was concomitant with the loss of a host cell. The transplanted and remaining host cells contained large vacuoles of ingested debris as well as lipofuscin-like granules, suggesting that they had scavenged the excess injected and host cells, and were stressed by the high digestive load. Therefore, although significant functional and structural recovery was observed, the consequences of this digestive stress may be a concern for longer-term health, especially where RPE cell transplantation is used to treat diseases that include lipofuscin accumulation as part of their pathology.

Keywords

RetinaSpectral domain optical coherence tomographyCell transplantationLipofuscinElectroretinography
Type
Research Articles
Information
Visual Neuroscience , Volume 29 , Issue 2 , March 2012 , pp. 83 - 93
Copyright
Copyright © Cambridge University Press 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anderson, D.H., Guerin, C.J., Erickson, P.A., Stern, W.H. & Fisher, S.K. (1986). Morphological recovery in the reattached retina. Investigative Ophthalmology & Visual Science 27, 168–183.Google ScholarPubMed
Anderson, D.H., Stern, W.H., Fisher, S.K., Erickson, P.A. & Borgula, G.A. (1981). The onset of pigment epithelial proliferation after retinal detachment. Investigative Ophthalmology & Visual Science 21, 10–16.Google ScholarPubMed
Anderson, D.H., Stern, W.H., Fisher, S.K., Erickson, P.A. & Borgula, G.A. (1983). Retinal detachment in the cat: The pigment epithelial-photoreceptor interface. Investigative Ophthalmology & Visual Science 24, 906–926.Google ScholarPubMed
Arai, S., Thomas, B.B., Seiler, M.J., Aramant, R.B., Qiu, G., Mui, C., de Juan, E. & Sadda, S.R. (2004). Restoration of visual responses following transplantation of intact retinal sheets in rd mice. Experimental Eye Research 79, 331–341.CrossRefGoogle ScholarPubMed
Cebulla, C.M., Ruggeri, M., Murray, T.G., Feuer, W.J. & Hernandez, E. (2010). Spectral domain optical coherence tomography in a murine retinal detachment model. Experimental Eye Research 90, 521–527.Google Scholar
Fischer, M.D., Huber, G., Beck, S.C., Tanimoto, N., Muehlfriedel, R., Fahl, E., Grimm, C., Wenzel, A., Reme, C.E., van de Pavert, S.A., Wijnholds, J., Pacal, M., Bremner, R. & Seeliger, M.W. (2009). Noninvasive, in vivo assessment of mouse retinal structure using optical coherence tomography. PLoS One 4, e7507.CrossRefGoogle ScholarPubMed
Futter, C.E., Ramalho, J.S., Jaissle, G.B., Seeliger, M.W. & Seabra, M.C. (2004). The role of Rab27a in the regulation of melanosome distribution within retinal pigment epithelial cells. Molecular Biology of the Cell 15, 2264–2275.CrossRefGoogle ScholarPubMed
Gabriele, M.L., Ishikawa, H., Schuman, J.S., Bilonick, R.A., Kim, J.S., Kagemann, L. & Wollstein, G. (2010). Reproducibility of spectral-domain optical coherence tomography total retinal thickness measurements in mice. Investigative Ophthalmology & Visual Science 51 (12): 6519–6523.CrossRefGoogle ScholarPubMed
Gibbs, D., Azarian, S.M., Lillo, C., Kitamoto, J., Klomp, A.E., Steel, K.P., Libby, R.T. & Williams, D.S. (2004). Role of myosin VIIa and Rab27a in the motility and localization of RPE melanosomes. Journal of Cell Science 117, 6473–6483.Google ScholarPubMed
Gibbs, D. & Williams, D.S. (2003). Isolation and culture of primary mouse retinal pigmented epithelial cells. Advances in Experimental Medicine & Biology 533, 347–352.CrossRefGoogle ScholarPubMed
Gouras, P., Kong, J. & Tsang, S.H. (2002). Retinal degeneration and RPE transplantation in Rpe65(−/−) mice. Investigative Ophthalmology & Visual Science 43, 3307–3311.Google ScholarPubMed
Huber, G., Beck, S.C., Grimm, C., Sahaboglu-Tekgoz, A., Paquet-Durand, F., Wenzel, A., Humphries, P., Redmond, T.M., Seeliger, M.W. & Fischer, M.D. (2009). Spectral domain optical coherence tomography in mouse models of retinal degeneration. Investigative Ophthalmology & Visual Science 50, 5888–5895.CrossRefGoogle ScholarPubMed
Lamba, D.A., Karl, M.O. & Reh, T.A. (2009). Strategies for retinal repair: Cell replacement and regeneration. Progress in Brain Research 175, 23–31.CrossRefGoogle ScholarPubMed
Li, L.X. & Turner, J.E. (1988 a). Transplantation of retinal pigment epithelial cells to immature and adult rat hosts: Short- and long-term survival characteristics. Experimental Eye Research 47, 771–785.CrossRefGoogle Scholar
Li, L.X. & Turner, J.E. (1988 b). Transplantation of retinal pigment epithelial cells to immature and adult rat hosts: Short- and long-term survival characteristics. Experimental Eye Research 47, 771–785.CrossRefGoogle Scholar
Li, L. & Turner, J.E. (1991). Optimal conditions for long-term photoreceptor cell rescue in RCS rats: The necessity for healthy RPE transplants. Experimental Eye Research 52, 669–679.CrossRefGoogle ScholarPubMed
Liu, X., Ondek, B. & Williams, D.S. (1998). Mutant myosin VIIa causes defective melanosome distribution in the RPE of shaker-1 mice. Nature Genetics 19, 117–118.CrossRefGoogle ScholarPubMed
Lopez, R., Gouras, P., Brittis, M. & Kjeldbye, H. (1987). Transplantation of cultured rabbit retinal epithelium to rabbit retina using a closed-eye method. Investigative Ophthalmology & Visual Science 28, 1131–1137.Google ScholarPubMed
Lu, B., Malcuit, C., Wang, S., Girman, S., Francis, P., Lemieux, L., Lanza, R. & Lund, R. (2009). Long-term safety and function of RPE from human embryonic stem cells in preclinical models of macular degeneration. Stem Cells 27, 2126–2135.CrossRefGoogle ScholarPubMed
Lund, R.D., Adamson, P., Sauve, Y., Keegan, D.J., Girman, S.V., Wang, S., Winton, H., Kanuga, N., Kwan, A.S., Beauchene, L., Zerbib, A., Hetherington, L., Couraud, P.O., Coffey, P. & Greenwood, J. (2001). Subretinal transplantation of genetically modified human cell lines attenuates loss of visual function in dystrophic rats. Proceedings of the National Academy of Sciences of the United States of America 98, 9942–9947.Google Scholar
Mata, N.L., Radu, R.A., Clemmons, R.C. & Travis, G.H. (2002). Isomerization and oxidation of vitamin a in cone-dominant retinas: A novel pathway for visual-pigment regeneration in daylight. Neuron 36, 69–80.CrossRefGoogle ScholarPubMed
Nusinowitz, S., Ridder, W.H. III. & Ramirez, J. (2007). Temporal response properties of the primary and secondary rod-signaling pathways in normal and Gnat2 mutant mice. Experimental Eye Research 84, 1104–1114.CrossRefGoogle ScholarPubMed
Pinilla, I., Lund, R.D., Lu, B. & Sauve, Y. (2005). Measuring the cone contribution to the ERG b-wave to assess function and predict anatomical rescue in RCS rats. Vision Research 45, 635–641.CrossRefGoogle Scholar
Ruggeri, M., Wehbe, H., Jiao, S., Gregori, G., Jockovich, M.E., Hackam, A., Duan, Y. & Puliafito, C.A. (2007). In vivo three-dimensional high-resolution imaging of rodent retina with spectral-domain optical coherence tomography. Investigative Ophthalmology & Visual Science 48, 1808–1814.CrossRefGoogle ScholarPubMed
Seiler, M.J., Rao, B., Aramant, R.B., Yu, L., Wang, Q., Kitayama, E., Pham, S., Yan, F., Chen, Z. & Keirstead, H.S. (2010). Three-dimensional optical coherence tomography imaging of retinal sheet implants in live rats. Journal of Neuroscience Methods 188, 250–257.CrossRefGoogle ScholarPubMed
Sheedlo, H.J., Li, L.X. & Turner, J.E. (1989). Functional and structural characteristics of photoreceptor cells rescued in RPE-cell grafted retinas of RCS dystrophic rats. Experimental Eye Research 48, 841–854.Google Scholar
Sheedlo, H.J., Li, L. & Turner, J.E. (1991). Photoreceptor cell rescue at early and late RPE-cell transplantation periods during retinal disease in RCS dystrophic rats. Journal of Neural Transplantation & Plasticity 2, 55–63.CrossRefGoogle ScholarPubMed
Timmers, A.M., Zhang, H., Squitieri, A. & Gonzalez-Pola, C. (2001). Subretinal injections in rodent eyes: Effects on electrophysiology and histology of rat retina. Molecular Vision 7, 131–137.Google ScholarPubMed
Weng, J., Mata, N.L., Azarian, S.M., Tzekov, R.T., Birch, D.G. & Travis, G.H. (1999). Insights into the function of Rim protein in photoreceptors and etiology of Stargardt’s disease from the phenotype in abcr knockout mice. Cell 98, 13–23.CrossRefGoogle ScholarPubMed
Zhang, X. & Bok, D. (1998). Transplantation of retinal pigment epithelial cells and immune response in the subretinal space. Investigative Ophthalmology & Visual Science 39, 1021–1027.Google ScholarPubMed