Guo Qiang, Zeng Yu-Xiao, Huang Shu-Dong, Zou Ting, Yin Zheng-Qin
Southwest Hospital/Southwest Eye Hospital, Third Military Medical University (Army Medical University), Chongqing 400038, China.
Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing 400038, China.
Int J Ophthalmol. 2023 Apr 18;16(4):483-498. doi: 10.18240/ijo.2023.04.01. eCollection 2023.
To explore whether the subretinal transplantation of retinal progenitor cells from human embryonic stem cell-derived retinal organoid (hERO-RPCs) could promote Müller glia dedifferentiation and transdifferentiation, thus improving visual function and delaying retinal degenerative progression.
hERO-RPCs were subretinally transplanted into Royal College of Surgeons (RCS) rats. Electroretinography (ERG) recording was performed at 4 and 8wk postoperation to assess retinal function. Using immunofluorescence, the changes in outer nuclear layer (ONL) thickness and retinal Müller glia were explored at 2, 4, and 8wk postoperation. To verify the effect of hERO-RPCs on Müller glia , we cocultured hERO-RPCs with Müller glia with a Transwell system. After coculture, Ki67 staining and quantitative polymerase chain reaction (qPCR) were performed to measure the proliferation and mRNA levels of Müller glia respectively. Cell migration experiment was used to detect the effect of hERO-RPCs on Müller glial migration. Comparisons between two groups were performed by the unpaired Student's -test, and comparisons among multiple groups were made with one-way ANOVA followed by Tukey's multiple comparison test.
The visual function and ONL thickness of RCS rats were significantly improved by transplantation of hERO-RPCs at 4 and 8wk postoperation. In addition to inhibiting gliosis at 4 and 8wk postoperation, hERO-RPCs significantly increased the expression of dedifferentiation-associated transcriptional factor in Müller glia and promoted the migration at 2, 4 and 8wk postoperation, but not the transdifferentiation of these cells in RCS rats. , using the Transwell system, we found that hERO-RPCs promoted the proliferation and migration of primary rat Müller glia and induced their dedifferentiation at the mRNA level.
These results show that hERO-RPCs might promote early dedifferentiation of Müller glia, which may provide novel insights into the mechanisms of stem cell therapy and Müller glial reprogramming, contributing to the development of novel therapies for retinal degeneration disorders.
探讨将人胚胎干细胞来源的视网膜类器官中的视网膜祖细胞（hERO-RPCs）进行视网膜下移植是否能促进Müller胶质细胞去分化和转分化，从而改善视觉功能并延缓视网膜退行性病变进程。
将hERO-RPCs视网膜下移植到皇家外科学院（RCS）大鼠体内。在术后4周和8周进行视网膜电图（ERG）记录以评估视网膜功能。利用免疫荧光技术，在术后2周、4周和8周探究外核层（ONL）厚度及视网膜Müller胶质细胞的变化。为验证hERO-RPCs对Müller胶质细胞的作用，我们使用Transwell系统将hERO-RPCs与Müller胶质细胞进行共培养。共培养后，分别进行Ki67染色和定量聚合酶链反应（qPCR）以检测Müller胶质细胞的增殖及mRNA水平。采用细胞迁移实验检测hERO-RPCs对Müller胶质细胞迁移的影响。两组间比较采用非配对学生t检验，多组间比较采用单因素方差分析，随后进行Tukey多重比较检验。
术后4周和8周，hERO-RPCs移植显著改善了RCS大鼠的视觉功能和ONL厚度。hERO-RPCs除了在术后4周和8周抑制胶质细胞增生外，还显著增加了RCS大鼠Müller胶质细胞中去分化相关转录因子的表达，并在术后2周、4周和8周促进了其迁移，但未促进这些细胞的转分化。利用Transwell系统，我们发现hERO-RPCs促进了原代大鼠Müller胶质细胞的增殖和迁移，并在mRNA水平诱导其去分化。
这些结果表明，hERO-RPCs可能促进Müller胶质细胞的早期去分化，这可能为干细胞治疗和Müller胶质细胞重编程机制提供新的见解，有助于开发视网膜退行性疾病的新疗法。
