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A new immunodeficient pigmented retinal degenerate rat strain to study transplantation of human cells without immunosuppression

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Abstract

Purpose

The goal of this study was to develop an immunodeficient rat model of retinal degeneration (RD nude rats) that will not reject transplanted human cells.

Methods

SD-Tg(S334ter)3Lav females homozygous for a mutated mouse rhodopsin transgene were mated with NTac:NIH-Whn (NIH nude) males homozygous for the Foxn1 rnu allele. Through selective breeding, a new stock, SD-Foxn1 Tg(S334ter)3Lav (RD nude) was generated such that all animals were homozygous for the Foxn1 rnu allele and either homo- or hemizygous for the S334ter transgene. PCR-based assays for both the Foxn1 rnu mutation and the S334ter transgene were developed for accurate genotyping. Immunodeficiency was tested by transplanting sheets of hESC-derived neural progenitor cells to the subretinal space of RD nude rats, and, as a control, NIH nude rats. Rats were killed between 8 and 184 days after surgery, and eye sections were analyzed for human, neuronal, and glial markers.

Results

After transplantation to RD nude and to NIH nude rats, hESC-derived neural progenitor cells differentiated to neuronal and glial cells, and migrated extensively from the transplant sheets throughout the host retina. Migration was more extensive in RD nude than in NIH nude rats. Already 8 days after transplantation, donor neuronal processes were found in the host inner plexiform layer. In addition, host glial cells extended processes into the transplants. The host retina showed the same photoreceptor degeneration pattern as in the immunocompetent SD-Tg(S334ter)3Lav rats. Recipients survived well after surgery.

Conclusions

This new rat model is useful for testing the effect of human cell transplantation on the restoration of vision without interference of immunosuppression.

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Abbreviations

hESC:

Human embryonic stem cells

RD:

Retinal degeneration

GC:

Ganglion cell layer

IP:

Inner plexiform layer

IN:

Inner nuclear layer

ON:

Outer nuclear layer

RPE:

Retinal pigment epithelium

H:

Host

T:

Transplant

CRALBP:

Cellular retinaldehyde binding protein

GFAP:

Glial fibrillary acidic protein

NF:

Neurofilament

MAP:

Microtubule-associated protein

NRL:

Neural retina-specific leucine zipper protein

Otx2:

Orthodenticle homolog 2 homeobox protein

PKC:

Protein kinase C

RA:

Retinoic acid

HA:

Hyaluronic acid

IST:

Insulin/Selenite/Transferrin

T3:

Triiodothyronine

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Acknowledgments

This work was supported by the Lincy Foundation (MJS, HSK); NIH P40OD011062 (ECB). Access to the Optical Biology Core facility of the Developmental Biology Center, a Shared Resource, was supported in part by a Cancer Center Support Grant (CA-62203) and a Center for Complex Biological Systems Support Grant (GM-076516) at the University of California, Irvine. MKJ was supported by CIRM TB1-1182. We thank Matthew M. LaVail, UCSF, for the founder breeding pairs of transgenic S334ter line 3 rats. The initial breeding of the strain was done at Taconic Inc., Albany, NY, USA. Personnel of Taconic involved in the project included Kaitlyn Waterbury, Jacob Luft, and Kimberly Meagher. We thank Gabriel Nistor (now California Stem Cells Inc., Irvine, CA, USA) for designing the hESC differentiation protocol and preparing the cells for transplantation.

Conflict of interest

MJS and RBA have proprietary interests in the implantation instrument and procedure (Ocular Transplantation LLC; patents #5,941,250; 6,159,218; 6,156,042); RBA is also an employee of Ocular Transplantation LLC. HSK was on the scientific advisory board of California Stem Cell, Inc. at the time of the experiments; and is now President and Chief Executive Officer of California Stem Cell, Inc. since December 2013.

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Authors and Affiliations

  1. Anatomy & Neurobiology/Reeve-Irvine Research Center, University of California, Irvine, CA, USA

    Magdalene J. Seiler, Robert B. Aramant, Melissa K. Jones, Dave L. Ferguson & Hans S. Keirstead

  2. Department of Biomedical Sciences/Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA, USA

    Melissa K. Jones

  3. Department of Biological Sciences, California State University Long Beach, Long Beach, CA, USA

    Melissa K. Jones

  4. Rat Resource and Research Center, University of Missouri, Columbia, MO, USA

    Elizabeth C. Bryda

  5. Reeve-Irvine Research Center, Sue and Bill Gross Stem Cell Research Center, School of Medicine, University of California at Irvine, Irvine, CA, USA

    Hans S. Keirstead

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  1. Magdalene J. Seiler
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Correspondence to Hans S. Keirstead.

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Seiler, M.J., Aramant, R.B., Jones, M.K. et al. A new immunodeficient pigmented retinal degenerate rat strain to study transplantation of human cells without immunosuppression. Graefes Arch Clin Exp Ophthalmol 252, 1079–1092 (2014). https://doi.org/10.1007/s00417-014-2638-y

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