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视网膜色素上皮萎缩的实验性干细胞替代移植

Experimental stem cell replacement transplantation in retinal pigment epithelium atrophy.

作者信息

Neroeva N V, Neroev V V, Katargina L A, Ryabina M V, Ilyukhin P A, Karmokova A G, Losanova O A, Maybogin A M, Kharitonov A E, Eremeev A V, Lagarkova M A

机构信息

Helmholtz National Medical Research Center of Eye Diseases, Moscow, Russia.

Federal Research and Clinical Center of Physical-Chemical Medicine, Moscow, Russia.

出版信息

Vestn Oftalmol. 2022;138(3):7-15. doi: 10.17116/oftalma20221380317.

Abstract

PURPOSE

To develop and evaluate the results of the modified surgical technique for transplantation of retinal pigment epithelium (RPE) differentiated from human induced pluripotent stem cells (iPSC-RPE) in the form of a cell suspension into the subretinal space of rabbits with previously induced RPE atrophy.

MATERIAL AND METHODS

The study was conducted on 10 New Zealand albino rabbits (20 eyes). One month after modeling RPE atrophy and retinal degeneration, rabbits were subjected to subretinal transplantation of iPSC-RPE cells in the form of a cell suspension. To prevent reflux of iPSC-RPE into the vitreal cavity, the injection site was sealed with 2-3 drops of autologous platelet-rich plasma (PRP). All rabbits underwent spectral optical coherence tomography (SOCT) and autofluorescence studies on the Heidelberg Spectralis system («Heidelberg Engineering», Germany). Enucleated animal eyes were studied with morphological and immunohistochemical methods.

RESULTS

In this study we developed and evaluated a modified surgical technique of transplantation of iPSC-RPE in the form of a cell suspension into the subretinal space of rabbits with induced RPE atrophy. It was found that the use of PRP helps seal the defect and prevents cell suspension reflux into the vitreous cavity, effectively minimizing intra- and postoperative complications. Morphological study and examination of histological sections showed that implantable iPSC-RPEs were correctly integrated and adhered to the choroid in the surgery site. Immunohistochemical analysis involving fluorescence-marked antibodies confirmed the survival of iPSC-RPE integrated into the retina of model animals.

CONCLUSION

This method improves the technology of iPSC-RPE transplantation on preclinical stages of the study, revealing new prospects in the treatment of degenerative diseases of the retina and the possibility of a personalized approach.

摘要

目的

开发并评估一种改良手术技术的效果,该技术是将从人诱导多能干细胞分化而来的视网膜色素上皮(iPSC-RPE)以细胞悬液的形式移植到先前诱导发生RPE萎缩的兔眼视网膜下间隙。

材料与方法

对10只新西兰白兔(20只眼)进行研究。在建立RPE萎缩和视网膜变性模型1个月后,将iPSC-RPE细胞以细胞悬液的形式进行视网膜下移植。为防止iPSC-RPE反流至玻璃体腔,注射部位用2 - 3滴自体富血小板血浆(PRP)封闭。所有兔子均在德国海德堡工程公司的海德堡Spectralis系统上进行光谱光学相干断层扫描(SOCT)和自发荧光研究。对摘除的动物眼球进行形态学和免疫组织化学研究。

结果

在本研究中,我们开发并评估了一种改良手术技术,即将iPSC-RPE以细胞悬液的形式移植到诱导发生RPE萎缩的兔眼视网膜下间隙。结果发现,使用PRP有助于封闭缺损并防止细胞悬液反流至玻璃体腔,有效减少术中及术后并发症。形态学研究和组织切片检查表明,可植入的iPSC-RPE在手术部位正确整合并附着于脉络膜。涉及荧光标记抗体的免疫组织化学分析证实,整合到模型动物视网膜中的iPSC-RPE存活。

结论

该方法改进了iPSC-RPE移植在研究临床前阶段的技术,为视网膜退行性疾病的治疗揭示了新的前景以及个性化治疗方法的可能性。

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