通过破坏神经胶质屏障增强移植的视网膜祖细胞向宿主视网膜的迁移。

Enhanced migration of engrafted retinal progenitor cells into the host retina via disruption of glial barriers.

机构信息

Eye Center, Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.

Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, China.

出版信息

Mol Vis. 2021 May 10;27:300-308. eCollection 2021.

PMID:34035644

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8131176/

Abstract

PURPOSE

Migration and integration remain critical challenges for stem cell replacement therapy. Glial barriers play an important role in preventing cell migration and integration. The purpose of this study was to investigate the effect and mechanisms of chondroitinase ABC on the migration of murine retinal progenitor cells (mRPCs) transplanted into the subretinal space of B6 mice.

METHODS

mRPCs were harvested from the neural retinas of P1 enhanced green fluorescent protein (GFP) B6 mice. Two μl containing 2 × 10 expanded RPCs alone or combined with chondroitinase ABC in suspension were injected into the subretinal space of the recipient B6 mice. Immunohistochemistry was performed on the recipient B6 retinas to evaluate the glial barrier formation and migration of the mRPCs. Western blotting was also used to check the expression of the glial barriers.

RESULTS

Glial fibrillary acidic protein (GFAP) and vimentin could be seen around the transplanted mRPCs in the B6 mice. Formation of glial barriers prevented the migration of donor cells into the retinal layers. Chondroitinase ABC promoted the migration and survival rates of the engrafted retinal progenitor cells in the retinal layers of recipient B6 mice. Injection induced upregulation of GFAP, chondroitin, and CD44 expression. Chondroitinase ABC disrupted the glial barriers. The CD44 around the mRPCs was much lower in the chondroitinase group. However, the CD44 in the retinal layers was considerably higher in the chondroitinase group. With the employment of chondroitinase ABC, more cells migrated into the outer nuclear layer or inner nuclear layer. The chondroitin and CD44 expression decreased 3 weeks after transplantation in the chondroitinase ABC group.

CONCLUSIONS

Chondroitinase ABC degraded glial barriers and enhanced the migration of transplanted mouse retinal progenitor cells. Chondroitinase ABC may also have induced activation of the CD44 signaling pathway to exert the effect.

摘要

目的

迁移和整合仍然是干细胞替代治疗的关键挑战。神经胶质细胞屏障在防止细胞迁移和整合方面发挥着重要作用。本研究旨在探讨软骨素酶 ABC 对移植到 B6 小鼠视网膜下腔的鼠视网膜祖细胞（mRPC）迁移的影响及其机制。

方法

从 P1 增强型绿色荧光蛋白（GFP）B6 小鼠的神经视网膜中分离 mRPC。将 2 μl 含有 2×10 个扩增 RPC 的悬浮液单独或与软骨素酶 ABC 混合注入受体 B6 小鼠的视网膜下腔。对受体 B6 视网膜进行免疫组织化学染色，以评估 mRPC 的胶质屏障形成和迁移。还使用 Western blot 检查胶质屏障的表达。

结果

在 B6 小鼠中，可以在移植的 mRPC 周围看到神经胶质纤维酸性蛋白（GFAP）和波形蛋白。胶质屏障的形成阻止了供体细胞向视网膜层迁移。软骨素酶 ABC 促进了植入的视网膜祖细胞在受体 B6 小鼠视网膜层中的迁移和存活率。注射诱导 GFAP、软骨素和 CD44 表达上调。软骨素酶 ABC 破坏了胶质屏障。软骨素酶组 mRPC 周围的 CD44 明显较低。然而，软骨素酶组的视网膜层中的 CD44 要高得多。使用软骨素酶 ABC，更多的细胞迁移到外核层或内核层。在软骨素酶 ABC 组，移植后 3 周时软骨素和 CD44 的表达降低。