Cargando…

Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules

Propagating large amounts of human corneal stromal cells (hCSCs) in vitro while maintaining the physiological quality of their phenotypes is necessary for their application in cell therapy. Here, a novel medium to propagate hCSCs obtained from small incision lenticule extraction (SMILE)‐derived lent...

Descripción completa

Detalles Bibliográficos
Autores principales: Li, Shenyang, Cui, Zekai, Gu, Jianing, Wang, Yini, Tang, Shibo, Chen, Jiansu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7812260/
https://www.ncbi.nlm.nih.gov/pubmed/33342057
http://dx.doi.org/10.1111/jcmm.16189
_version_ 1783637632582942720
author Li, Shenyang
Cui, Zekai
Gu, Jianing
Wang, Yini
Tang, Shibo
Chen, Jiansu
author_facet Li, Shenyang
Cui, Zekai
Gu, Jianing
Wang, Yini
Tang, Shibo
Chen, Jiansu
author_sort Li, Shenyang
collection PubMed
description Propagating large amounts of human corneal stromal cells (hCSCs) in vitro while maintaining the physiological quality of their phenotypes is necessary for their application in cell therapy. Here, a novel medium to propagate hCSCs obtained from small incision lenticule extraction (SMILE)‐derived lenticules was investigated and the feasibility of intrastromal injection of these hCSCs was assessed. Primary hCSCs were cultured in porcine corneal stroma extract (pCSE) with RIFA medium including ROCK inhibitor Y27632, insulin‐transferrin‐selenium, fibroblast growth factor 2, L‐ascorbate 2‐phosphate and 0.5% FBS (RIFA medium + pCSE). Protein profiling of the pCSE was identified using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC‐MS/MS). After subculturing in RIFA medium + pCSE or 10% FBS normal medium (NM), hCSCs at P4 were transplanted into mouse corneal stroma. Compared with NM, ALDH3A1, keratocan and lumican were significantly more expressed in the RIFA medium + pCSE. ALDH3A1 was also more expressed in the RIFA medium + pCSE than in the RIFA medium. Fibronectin and α‐SMA were less expressed in the RIFA medium + pCSE than in the NM. Using Metascape analysis, the pCSE with its anti‐fibrosis, pro‐proliferation and anti‐apoptosis activities, was beneficial for hCSC cultivation. The intrastromally implanted hCSCs in the RIFA medium + pCSE had positive CD34 expression but negative CD45 expression 35 days after injection. We provide a valuable new medium that is advantageous for the proliferation of hCSCs with the properties of physiological keratocytes. Intrastromal injection of hCSCs in RIFA medium + pCSE has the potential for clinical cell therapy.
format Online
Article
Text
id pubmed-7812260
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-78122602021-01-22 Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules Li, Shenyang Cui, Zekai Gu, Jianing Wang, Yini Tang, Shibo Chen, Jiansu J Cell Mol Med Original Articles Propagating large amounts of human corneal stromal cells (hCSCs) in vitro while maintaining the physiological quality of their phenotypes is necessary for their application in cell therapy. Here, a novel medium to propagate hCSCs obtained from small incision lenticule extraction (SMILE)‐derived lenticules was investigated and the feasibility of intrastromal injection of these hCSCs was assessed. Primary hCSCs were cultured in porcine corneal stroma extract (pCSE) with RIFA medium including ROCK inhibitor Y27632, insulin‐transferrin‐selenium, fibroblast growth factor 2, L‐ascorbate 2‐phosphate and 0.5% FBS (RIFA medium + pCSE). Protein profiling of the pCSE was identified using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC‐MS/MS). After subculturing in RIFA medium + pCSE or 10% FBS normal medium (NM), hCSCs at P4 were transplanted into mouse corneal stroma. Compared with NM, ALDH3A1, keratocan and lumican were significantly more expressed in the RIFA medium + pCSE. ALDH3A1 was also more expressed in the RIFA medium + pCSE than in the RIFA medium. Fibronectin and α‐SMA were less expressed in the RIFA medium + pCSE than in the NM. Using Metascape analysis, the pCSE with its anti‐fibrosis, pro‐proliferation and anti‐apoptosis activities, was beneficial for hCSC cultivation. The intrastromally implanted hCSCs in the RIFA medium + pCSE had positive CD34 expression but negative CD45 expression 35 days after injection. We provide a valuable new medium that is advantageous for the proliferation of hCSCs with the properties of physiological keratocytes. Intrastromal injection of hCSCs in RIFA medium + pCSE has the potential for clinical cell therapy. John Wiley and Sons Inc. 2020-12-20 2021-01 /pmc/articles/PMC7812260/ /pubmed/33342057 http://dx.doi.org/10.1111/jcmm.16189 Text en © 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Li, Shenyang
Cui, Zekai
Gu, Jianing
Wang, Yini
Tang, Shibo
Chen, Jiansu
Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title_full Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title_fullStr Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title_full_unstemmed Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title_short Effect of porcine corneal stromal extract on keratocytes from SMILE‐derived lenticules
title_sort effect of porcine corneal stromal extract on keratocytes from smile‐derived lenticules
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7812260/
https://www.ncbi.nlm.nih.gov/pubmed/33342057
http://dx.doi.org/10.1111/jcmm.16189
work_keys_str_mv AT lishenyang effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules
AT cuizekai effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules
AT gujianing effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules
AT wangyini effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules
AT tangshibo effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules
AT chenjiansu effectofporcinecornealstromalextractonkeratocytesfromsmilederivedlenticules