TY - JOUR
T1 - Differentiation and Maturation Effect of All-Trans Retinoic Acid on Cultured Fetal RPE and Stem Cell-Derived RPE Cells for Cell-Based Therapy
AU - Yan, Tingyu
AU - Yang, Na
AU - Hu, Wei
AU - Zhang, Xinxin
AU - Li, Xuedong
AU - Wang, Youjin
AU - Kong, Jun
N1 - Funding Information:
This work was supported by Liaoning Guangfu Hereditary Eye Disease Philanthropic Foundation (No. #2900020001). We wish to thank Dr. Neil Bressler from John Hopkins University for reviewing the manuscript. We thank Prof. Tao Meng from the First Affiliated Hospital of China Medical University for helping with the tissue donation and Prof. Weidong Li from Shanghai Jiaotong University for providing cells.
Publisher Copyright:
© 2022 The Author(s). Published with license by Taylor & Francis Group, LLC.
PY - 2022
Y1 - 2022
N2 - Purpose: Clinical trials using fetal retinal pigment epithelium (fRPE), human embryonic stem cell (hESC)-derived RPE, or human induced pluripotent stem cell (hiPSC)-derived RPE for cell-based therapy for degenerative retinal diseases have been carried out. We investigated the culture-induced changes in passaged fRPE, hESC-RPE, and hiPSC-RPE cells and explored the differentiation and maturation effect of all-trans retinoic acid (ATRA) on cells for manufacturing and screening high-quality RPE cells for clinical transplantation. Methods: RPE cell lines were set up and the culture-induced changes in subsequent passages caused by manipulating plating density, dissociation method, and repeated passaging were studied by a microscope, real-time quantitative PCR, western blot, and immunofluorescent assays. Gene and protein expression and functional characteristics of RPE cells incubated with ATRA were evaluated. Results: Compared with fRPE, hESC-RPE, and hiPSC-RPE showed decreased gene and protein expression of RPE markers. RPE cells underwent mesenchymal changes showing increased expression of mesenchymal markers including a-SMA, N-cadherin, fibronectin and decreased expression of RPE markers including RPE65, E-cadherin, and ZO-1, as a subsequence of low plating density, inappropriate dissociated method, and repeated passaging. RPE cells treated by ATRA showed increased expression of RPE markers and increased expression of negative complement regulatory proteins (CRPs) and increased transepithelial resistance as well. Conclusions: Differences in protein and gene expression among three RPE types exist. ATRA can increase RPE markers, CRPs gene expression in fRPE, and stem cell-derived RPE. These can be used to guide the standard of screening RPE cells for clinical translational cell therapy.
AB - Purpose: Clinical trials using fetal retinal pigment epithelium (fRPE), human embryonic stem cell (hESC)-derived RPE, or human induced pluripotent stem cell (hiPSC)-derived RPE for cell-based therapy for degenerative retinal diseases have been carried out. We investigated the culture-induced changes in passaged fRPE, hESC-RPE, and hiPSC-RPE cells and explored the differentiation and maturation effect of all-trans retinoic acid (ATRA) on cells for manufacturing and screening high-quality RPE cells for clinical transplantation. Methods: RPE cell lines were set up and the culture-induced changes in subsequent passages caused by manipulating plating density, dissociation method, and repeated passaging were studied by a microscope, real-time quantitative PCR, western blot, and immunofluorescent assays. Gene and protein expression and functional characteristics of RPE cells incubated with ATRA were evaluated. Results: Compared with fRPE, hESC-RPE, and hiPSC-RPE showed decreased gene and protein expression of RPE markers. RPE cells underwent mesenchymal changes showing increased expression of mesenchymal markers including a-SMA, N-cadherin, fibronectin and decreased expression of RPE markers including RPE65, E-cadherin, and ZO-1, as a subsequence of low plating density, inappropriate dissociated method, and repeated passaging. RPE cells treated by ATRA showed increased expression of RPE markers and increased expression of negative complement regulatory proteins (CRPs) and increased transepithelial resistance as well. Conclusions: Differences in protein and gene expression among three RPE types exist. ATRA can increase RPE markers, CRPs gene expression in fRPE, and stem cell-derived RPE. These can be used to guide the standard of screening RPE cells for clinical translational cell therapy.
KW - Retinal pigment epithelium
KW - all-trans retinoic acid
KW - epithelial-mesenchymal transition
KW - human embryonic stem cells
KW - human induced pluripotent stem cells
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U2 - 10.1080/02713683.2022.2079144
DO - 10.1080/02713683.2022.2079144
M3 - Article
C2 - 35763026
AN - SCOPUS:85135113142
SN - 0271-3683
VL - 47
SP - 1300
EP - 1311
JO - Current Eye Research
JF - Current Eye Research
IS - 9
ER -