Senior Department of Ophthalmology, 3rd Medical Center of Chinese PLA General Hospital, Beijing, China.
State Key Laboratory of Stem Cell and Reproductive Biology, National Stem Cell Resource Center, Institute of Zoology, Chinese Academy of Sciences, Beijing, China; University of Chinese Academy of Sciences, Beijing, China.
Cytotherapy. 2024 Jun;26(6):606-615. doi: 10.1016/j.jcyt.2024.02.020. Epub 2024 Mar 4.
Mesenchymal stromal cells (MSCs) hold great promise in the treatment of diabetic retinopathy (DR), as evidenced by increasing preclinical and clinical studies. However, the absence of standardized and industrialized clinical-grade donor cells hampers the continued development and large-scale clinical application of MSCs-based therapies for DR. Previously, we have identified a unique population of MSCs generated from a clinical-grade human embryonic stem cell (hESC) line under Good Manufacturing Practice conditions that could be a potential source to address the issues. Here, we investigated the therapeutic potential of the clinical-grade hESC line-derived MSCs (hESC-MSCs) on db/db mice with DR.
hESC-MSCs were initially characterized by morphological assessment, flow cytometry analysis and trilineage differentiation assays. These cells (5 × 10 cells) were then transplanted intravenously into 12-week-old db/db mice via tail vein, with phosphate-buffered saline transplantation and untreated groups used as controls. The retinal alterations in neural functions and microvascular perfusions, and inflammatory responses in peripheral blood and retina were evaluated at 4 and 6 weeks after transplantation using electroretinography, optical coherence tomography angiography and flow cytometry, respectively. Body weight and fasting blood glucose (FBG) levels were also measured to investigate their systemic implications.
Compared with controls, intravenous transplantation of hESC-MSCs could significantly: (i) enhance impaired retinal electroretinography functions (including amplitudes of a-, b-wave and oscillatory potentials) at 4 weeks after transplantation; (ii) alleviate microvascular dysfunctions, especially in the inner retina with significance (including reducing non-perfusion area and increasing vascular area density) at 4 weeks after transplantation; (iii) decrease FBG levels at 4 weeks after transplantation and induce weight loss up to 6 weeks after transplantation and (iv) increase both peripheral blood and retinal interleukin-10 levels at 4 weeks after transplantation and modulate peripheral blood inflammatory cytokines and chemokines levels, such as monocyte chemotactic protein-1, up to 6 weeks after transplantation.
The findings of our study indicated that intravenous transplantation of hESC-MSCs ameliorated retinal neural and microvascular dysfunctions, regulated body weight and FBG and modulated peripheral blood and retinal inflammation responses in a mouse model of DR. These results suggest that hESC-MSCs could be a potentially effective clinical-grade cell source for the treatment of DR.
间充质基质细胞（MSCs）在治疗糖尿病视网膜病变（DR）方面具有巨大的潜力，越来越多的临床前和临床研究证明了这一点。然而，缺乏标准化和工业化的临床级供体细胞阻碍了基于 MSCs 的治疗 DR 的进一步发展和大规模临床应用。此前，我们已经从符合良好生产规范的临床级人胚胎干细胞（hESC）系中鉴定出一种独特的 MSC 群体，它们可能是解决这一问题的潜在来源。在这里，我们研究了临床级 hESC 系衍生的 MSC（hESC-MSCs）对 DR 模型 db/db 小鼠的治疗潜力。
首先通过形态评估、流式细胞术分析和三系分化实验对 hESC-MSCs 进行特征描述。然后，将这些细胞（5×10 个细胞）通过尾静脉静脉内移植到 12 周龄 db/db 小鼠体内，磷酸盐缓冲盐水移植和未处理组作为对照。在移植后 4 周和 6 周，分别通过视网膜电图、光学相干断层扫描血管造影和流式细胞术评估神经功能和微血管灌注的视网膜改变以及外周血和视网膜中的炎症反应。还测量了体重和空腹血糖（FBG）水平，以研究其全身影响。
与对照组相比，静脉内移植 hESC-MSCs 可以显著：（i）在移植后 4 周时增强受损的视网膜视网膜电图功能（包括 a-、b-波和振荡电位的幅度）；（ii）在移植后 4 周时缓解微血管功能障碍，特别是在内层视网膜（包括减少无灌注区和增加血管面积密度）；（iii）在移植后 4 周时降低 FBG 水平，并在移植后 6 周时引起体重减轻；（iv）在移植后 4 周时增加外周血和视网膜白细胞介素-10 水平，并在移植后 6 周时调节外周血炎症细胞因子和趋化因子水平，如单核细胞趋化蛋白-1。
我们的研究结果表明，静脉内移植 hESC-MSCs 可改善 DR 模型小鼠的视网膜神经和微血管功能障碍，调节体重和 FBG，并调节外周血和视网膜炎症反应。这些结果表明，hESC-MSCs 可能是治疗 DR 的一种潜在有效的临床级细胞来源。
