Momenzadeh Sara, Karamali Fereshteh, Atefi Atefeh, Nasr-Esfahani Mohammad Hossein
Higher Education Jahad University of Isfahan Province, Isfahan, Iran.
Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Cell J. 2022 Mar;24(3):127-132. doi: 10.22074/cellj.2022.7764.
Degeneration of the photoreceptors due to retinal disorders can affect vision, and even lead to blindness. Recently therapeutic progress in retinal degeneration, using human embryonic stem cells (hESCs), has been facing technical challenges, demanding the development of simple and standardized protocols. In addition to the designing of the protocols, characterization of the obtained cells is highly required for confirming the reliability of the applied methods for future medical applications. Previously, we showed that human stem cells from apical papilla (SCAP) have stromal cell-derived inducing activity (SDIA).
In this experimental study, we developed an efficient retinal differentiation protocol, based on the co-culture of confluent hESCs and SCAP in the absence of exogenous molecules, such as activators or inhibitors of molecular signaling pathways. This experimental procedure resulted in the generation of self-forming neural retina (NR)-like structures containing retinal progenitor cells (RPCs) within 4 weeks.
We have focused on the characterization of the derived RPCs, as a crucial step towards further verification of the efficiency of our previously suggested protocol. The differentiated cells expressed eye-field markers, PAX6, RAX, LHX2, and SIX3, and also generated neurospheres by a floating culture system for one week.
We have reported that the treatment of hESC-derived RPCs by the Notch pathway-inhibitor induced the generation of photoreceptor precursor cells (PPCs). The presented method demonstrates the fact that a co-culture of hESCs and SCAP without exogenous molecules provides an efficient approach to produce RPCs for the treatment of retinal disease, and act as an model for the development of human retina.
视网膜疾病导致的光感受器退化会影响视力，甚至导致失明。最近，利用人类胚胎干细胞（hESCs）治疗视网膜退化的进展面临技术挑战，需要开发简单且标准化的方案。除了设计方案外，为了确认所应用方法在未来医学应用中的可靠性，对获得的细胞进行表征也非常必要。此前，我们发现根尖乳头来源的人类干细胞（SCAP）具有基质细胞衍生诱导活性（SDIA）。
在本实验研究中，我们基于汇合的hESCs与SCAP在无外源分子（如分子信号通路激活剂或抑制剂）的情况下共培养，开发了一种高效的视网膜分化方案。该实验过程在4周内产生了包含视网膜祖细胞（RPCs）的自形成神经视网膜（NR）样结构。
我们专注于对所衍生的RPCs进行表征，这是进一步验证我们之前提出的方案效率的关键步骤。分化细胞表达眼场标记物PAX6、RAX、LHX2和SIX3，并且通过悬浮培养系统培养一周后还能生成神经球。
我们曾报道，通过Notch信号通路抑制剂处理hESC衍生的RPCs可诱导光感受器前体细胞（PPCs）的产生。本文所展示的方法表明，在无外源分子的情况下，hESCs与SCAP共培养为治疗视网膜疾病产生RPCs提供了一种有效的方法，并可作为人类视网膜发育的模型。
