A new immunodeficient pigmented retinal degenerate rat strain to study transplantation of human cells without immunosuppression
A new immunodeficient pigmented retinal degenerate rat strain to study transplantation of human cells without immunosuppression
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DOI：
10.1007/s00417-014-2638-y
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发表时间：
2014-07-01
影响因子：
2.7
通讯作者：
Keirstead, Hans S.
中科院分区：
文献类型：
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作者：
Seiler, Magdalene J.;Aramant, Robert B.;Keirstead, Hans S.
将突变小鼠视紫红质转基因的SD-Tg（S334 ter）3Lav纯合子雌性与Foxn 1（rnu）等位基因纯合子的NTac：NIH-Whn（NIH裸）雄性交配。通过选择性育种，产生了新的原种SD-Foxn 1 Tg（S334 ter）3Lav（RD裸），使得所有动物对于Foxn 1（rnu）等位基因是纯合的，并且对于S334 ter转基因是纯合或半合的。Foxn 1（rnu）突变和S334 ter转基因的PCR为基础的分析开发了准确的基因分型。通过将hESC衍生的神经前体细胞片移植到RD裸大鼠的视网膜下空间来测试免疫缺陷，并且作为对照，NIH裸大鼠。在手术后8 - 184天处死大鼠，分析眼切片中的人、神经元和神经胶质标记物。移植到RD裸大鼠和NIH裸大鼠后，hESC衍生的神经前体细胞分化为神经元和神经胶质细胞，并从移植片广泛迁移到整个宿主视网膜。RD裸大鼠的迁移比NIH裸大鼠更广泛。移植后8天，在宿主内丛状层中发现供体神经元突起。此外，宿主神经胶质细胞将突起延伸到移植物中。宿主视网膜表现出与免疫活性SD-Tg（S334 ter）3Lav大鼠相同的光感受器变性模式。这种新的大鼠模型可用于在不受免疫抑制干扰的情况下检测人细胞移植对视力恢复的影响。
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.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.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.This new rat model is useful for testing the effect of human cell transplantation on the restoration of vision without interference of immunosuppression.