Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet

Abstract Transplantation of cultivated limbal epithelial transplantation has been proven to restore the corneal surface in limbal stem cell deficiency (LSCD). Here we comparatively investigated the optimized conditions and the efficiency of limbal epithelial sheet growth in three media conditions as...

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Autores principales:	Hyun Jung Lee, Sang Min Nam, Sae Kyung Choi, Kyoung Yul Seo, Hyun Ok Kim, So-Hyang Chung
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	ATP-binding Cassette Sub-family G Member 2 (ABCG2) Limbal Stem Cell Deficiency (LSCD) Outgrowth Sheets Limbal Explants Low JC Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7bacb59a0fbd4f4e8a4bf83f5c937a5e
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spelling	oai:doaj.org-article:7bacb59a0fbd4f4e8a4bf83f5c937a5e2021-12-02T11:40:17ZComparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet10.1038/s41598-018-32914-02045-2322https://doaj.org/article/7bacb59a0fbd4f4e8a4bf83f5c937a5e2018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-32914-0https://doaj.org/toc/2045-2322Abstract Transplantation of cultivated limbal epithelial transplantation has been proven to restore the corneal surface in limbal stem cell deficiency (LSCD). Here we comparatively investigated the optimized conditions and the efficiency of limbal epithelial sheet growth in three media conditions as well as with substrate free (transwell), human amniotic membrane (HAM) sutured onto transwell inserts (HAMTW), and HAM slide scaffold (HAMS). Outcomes evaluated were outgrowth sheet size from limbal explants, expression of stem/progenitor cell markers p63α, ABCG2 and CK15, and colony formation efficiency (CFE). Additionally, limbal epithelial sheets on HAMS were transplanted into corneas of LSCD rabbit models. Limbal epithelial sheets with 5% human AB serum showed the greatest increase in ABCG2 efflux activity (JC1low), p63α expression, and CFE compared in both conditions without HAM and with HAM, respectively. The outgrowth sheet size, cell yield, and Ki67 expression were increased in limbal epithelial sheets on HAMS compared to transwell and HAMTW. ABCG2 efflux activity, p63α and CK15 expressions, and CFE were also increased in limbal epithelial sheets on HAMS as well. In corneas of transplanted rabbit LSCD models, p63α expressions were noted in the basal layers and CK12 expressions were observed in superficial layers. Cultivation of limbal epithelial sheet on HAMS with xeno-free medium enhances the growth and stemness of limbal epithelial sheets.Hyun Jung LeeSang Min NamSae Kyung ChoiKyoung Yul SeoHyun Ok KimSo-Hyang ChungNature PortfolioarticleATP-binding Cassette Sub-family G Member 2 (ABCG2)Limbal Stem Cell Deficiency (LSCD)Outgrowth SheetsLimbal ExplantsLow JCMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	ATP-binding Cassette Sub-family G Member 2 (ABCG2) Limbal Stem Cell Deficiency (LSCD) Outgrowth Sheets Limbal Explants Low JC Medicine R Science Q
spellingShingle	ATP-binding Cassette Sub-family G Member 2 (ABCG2) Limbal Stem Cell Deficiency (LSCD) Outgrowth Sheets Limbal Explants Low JC Medicine R Science Q Hyun Jung Lee Sang Min Nam Sae Kyung Choi Kyoung Yul Seo Hyun Ok Kim So-Hyang Chung Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
description	Abstract Transplantation of cultivated limbal epithelial transplantation has been proven to restore the corneal surface in limbal stem cell deficiency (LSCD). Here we comparatively investigated the optimized conditions and the efficiency of limbal epithelial sheet growth in three media conditions as well as with substrate free (transwell), human amniotic membrane (HAM) sutured onto transwell inserts (HAMTW), and HAM slide scaffold (HAMS). Outcomes evaluated were outgrowth sheet size from limbal explants, expression of stem/progenitor cell markers p63α, ABCG2 and CK15, and colony formation efficiency (CFE). Additionally, limbal epithelial sheets on HAMS were transplanted into corneas of LSCD rabbit models. Limbal epithelial sheets with 5% human AB serum showed the greatest increase in ABCG2 efflux activity (JC1low), p63α expression, and CFE compared in both conditions without HAM and with HAM, respectively. The outgrowth sheet size, cell yield, and Ki67 expression were increased in limbal epithelial sheets on HAMS compared to transwell and HAMTW. ABCG2 efflux activity, p63α and CK15 expressions, and CFE were also increased in limbal epithelial sheets on HAMS as well. In corneas of transplanted rabbit LSCD models, p63α expressions were noted in the basal layers and CK12 expressions were observed in superficial layers. Cultivation of limbal epithelial sheet on HAMS with xeno-free medium enhances the growth and stemness of limbal epithelial sheets.
format	article
author	Hyun Jung Lee Sang Min Nam Sae Kyung Choi Kyoung Yul Seo Hyun Ok Kim So-Hyang Chung
author_facet	Hyun Jung Lee Sang Min Nam Sae Kyung Choi Kyoung Yul Seo Hyun Ok Kim So-Hyang Chung
author_sort	Hyun Jung Lee
title	Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
title_short	Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
title_full	Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
title_fullStr	Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
title_full_unstemmed	Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
title_sort	comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/7bacb59a0fbd4f4e8a4bf83f5c937a5e
work_keys_str_mv	AT hyunjunglee comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet AT sangminnam comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet AT saekyungchoi comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet AT kyoungyulseo comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet AT hyunokkim comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet AT sohyangchung comparativestudyofsubstratefreeandamnioticmembranescaffoldsforcultivationoflimbalepithelialsheet
_version_	1718395653749473280

Comparative study of substrate free and amniotic membrane scaffolds for cultivation of limbal epithelial sheet

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