학술논문
Formation of three-dimensional cell aggregates expressing lens-specific proteins in various cultures of human iris-derived tissue cells and iPS cells
induced pluripotent stem cells
induced pluripotent stem cells
Document Type
Report
Author
Source
Experimental and Therapeutic Medicine. August 2022, Vol. 24 Issue 2
Subject
Language
English
ISSN
1792-0981
Abstract
Introduction The development of the lens in the eye begins with the PAX6-expressing epidermal ectoderm making contact with the optic vesicle. Subsequently, SOX2 is expressed in the ectoderm region in [...]
Induced pluripotent stem (iPS) cells are widely used as a research tool in regenerative medicine and embryology. In studies related to lens regeneration in the eye, iPS cells have been reported to differentiate into lens epithelial cells (LECs); however, to the best of our knowledge, no study to date has described their formation of three-dimensional cell aggregates. Notably, in vivo studies in newts have revealed that iris cells in the eye can dedifferentiate into LECs and regenerate a new lens. Thus, as basic research on lens regeneration, the present study investigated the differentiation of human iris tissue-derived cells and human iris tissue-derived iPS cells into LECs and their formation of three-dimensional cell aggregates using a combination of two-dimensional culture, static suspension culture and rotational suspension culture. The results revealed that three-dimensional cell aggregates were formed and differentiated into LECs expressing [alpha]A-crystallin, a specific marker protein for LECs, suggesting that the cell-cell interaction facilitated by cell aggregation may have a critical role in enabling highly efficient differentiation of LECs. However, the present study was unable to achieve transparency in the cell aggregates; therefore, we aim to continue to investigate the degradation of organelles and other materials necessary to make the interior of the formed cell aggregates transparent. Furthermore, we aim to expand on our current work to study the regeneration of the lens and ciliary body as a whole in vitro, with the aim of being able to restore focusing function after cataract surgery. Abbreviations: DMEM/F12, Dulbecco's modified Eagle's medium/Ham's F12; b-FGF, basic fibroblastic growth factor; ES cells, embryonic stem cells; H&E, hematoxylin and eosin; H-iris cells, human iris tissue-derived cells; H-iris iPS cells, human iris-derived iPS cells; IOLs, intraocular lenses; iPS cells, induced pluripotent stem cells; KSR, KnockOut[TM] serum replacement; LECs, lens epithelial cells; LFCs, lens fiber cells; PECs, pigmented epithelial cells; qPCR, quantitative PCR; SEAM, self-formed ectodermal autonomous multi-zone Key words: iPS cells, LECs, H-iris cells, three-dimensional cell aggregate, p75NTR, [alpha]A-crystallin, type IV collagen, SEAM
Induced pluripotent stem (iPS) cells are widely used as a research tool in regenerative medicine and embryology. In studies related to lens regeneration in the eye, iPS cells have been reported to differentiate into lens epithelial cells (LECs); however, to the best of our knowledge, no study to date has described their formation of three-dimensional cell aggregates. Notably, in vivo studies in newts have revealed that iris cells in the eye can dedifferentiate into LECs and regenerate a new lens. Thus, as basic research on lens regeneration, the present study investigated the differentiation of human iris tissue-derived cells and human iris tissue-derived iPS cells into LECs and their formation of three-dimensional cell aggregates using a combination of two-dimensional culture, static suspension culture and rotational suspension culture. The results revealed that three-dimensional cell aggregates were formed and differentiated into LECs expressing [alpha]A-crystallin, a specific marker protein for LECs, suggesting that the cell-cell interaction facilitated by cell aggregation may have a critical role in enabling highly efficient differentiation of LECs. However, the present study was unable to achieve transparency in the cell aggregates; therefore, we aim to continue to investigate the degradation of organelles and other materials necessary to make the interior of the formed cell aggregates transparent. Furthermore, we aim to expand on our current work to study the regeneration of the lens and ciliary body as a whole in vitro, with the aim of being able to restore focusing function after cataract surgery. Abbreviations: DMEM/F12, Dulbecco's modified Eagle's medium/Ham's F12; b-FGF, basic fibroblastic growth factor; ES cells, embryonic stem cells; H&E, hematoxylin and eosin; H-iris cells, human iris tissue-derived cells; H-iris iPS cells, human iris-derived iPS cells; IOLs, intraocular lenses; iPS cells, induced pluripotent stem cells; KSR, KnockOut[TM] serum replacement; LECs, lens epithelial cells; LFCs, lens fiber cells; PECs, pigmented epithelial cells; qPCR, quantitative PCR; SEAM, self-formed ectodermal autonomous multi-zone Key words: iPS cells, LECs, H-iris cells, three-dimensional cell aggregate, p75NTR, [alpha]A-crystallin, type IV collagen, SEAM