Isolation and characterization of novel primary cells from the human distal outflow pathway

Abstract Ocular hypertension occurs due to increased resistance to aqueous humor removal through the conventional outflow pathway. Unlike the proximal region of the conventional outflow pathway, the distal region has not been well studied, mostly due to lack of model systems. Here we describe isolat...

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Autores principales:	Uttio Roy Chowdhury, Cindy K. Bahler, Cheryl R. Hann, Bradley H. Holman, Michael P. Fautsch
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1cf5c2287ff2434d90daf7456fc0bb71
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spelling	oai:doaj.org-article:1cf5c2287ff2434d90daf7456fc0bb712021-12-02T10:54:06ZIsolation and characterization of novel primary cells from the human distal outflow pathway10.1038/s41598-021-83558-62045-2322https://doaj.org/article/1cf5c2287ff2434d90daf7456fc0bb712021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83558-6https://doaj.org/toc/2045-2322Abstract Ocular hypertension occurs due to increased resistance to aqueous humor removal through the conventional outflow pathway. Unlike the proximal region of the conventional outflow pathway, the distal region has not been well studied, mostly due to lack of model systems. Here we describe isolation and characterization of human primary vascular distal outflow pathway (VDOP) cells from the distal region of the conventional outflow pathway. Tissue from the distal region was isolated from human corneo-scleral rims, digested with collagenase type I (100 U/ml) and placed on gelatin coated plates to allow cellular growth in Dulbecco’s Modified Eagle’s Medium (low glucose) containing fetal bovine serum and antibiotic/antimycotic. VDOP cells showed consistent proliferation for up to 7 passages, retained endothelial-like nature of the parent tissues and showed a unique marker phenotype of Lectin+VEGFR2-CD34-NG2- that was distinct from neighboring trabecular meshwork (Lectin+VEGFR2-CD34-NG2+) and Schlemm’s canal (Lectin+VEGFR2+CD34+NG2+) cells. Dexamethasone treated VDOP cells did not express myocilin and did not form cross-linked actin networks, in contrast to trabecular meshwork cells. These data show that VDOP cells are unique to the distal outflow region and can be used as a viable in vitro model system to understand the biology of the distal outflow pathway and intraocular pressure regulation.Uttio Roy ChowdhuryCindy K. BahlerCheryl R. HannBradley H. HolmanMichael P. FautschNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Uttio Roy Chowdhury Cindy K. Bahler Cheryl R. Hann Bradley H. Holman Michael P. Fautsch Isolation and characterization of novel primary cells from the human distal outflow pathway
description	Abstract Ocular hypertension occurs due to increased resistance to aqueous humor removal through the conventional outflow pathway. Unlike the proximal region of the conventional outflow pathway, the distal region has not been well studied, mostly due to lack of model systems. Here we describe isolation and characterization of human primary vascular distal outflow pathway (VDOP) cells from the distal region of the conventional outflow pathway. Tissue from the distal region was isolated from human corneo-scleral rims, digested with collagenase type I (100 U/ml) and placed on gelatin coated plates to allow cellular growth in Dulbecco’s Modified Eagle’s Medium (low glucose) containing fetal bovine serum and antibiotic/antimycotic. VDOP cells showed consistent proliferation for up to 7 passages, retained endothelial-like nature of the parent tissues and showed a unique marker phenotype of Lectin+VEGFR2-CD34-NG2- that was distinct from neighboring trabecular meshwork (Lectin+VEGFR2-CD34-NG2+) and Schlemm’s canal (Lectin+VEGFR2+CD34+NG2+) cells. Dexamethasone treated VDOP cells did not express myocilin and did not form cross-linked actin networks, in contrast to trabecular meshwork cells. These data show that VDOP cells are unique to the distal outflow region and can be used as a viable in vitro model system to understand the biology of the distal outflow pathway and intraocular pressure regulation.
format	article
author	Uttio Roy Chowdhury Cindy K. Bahler Cheryl R. Hann Bradley H. Holman Michael P. Fautsch
author_facet	Uttio Roy Chowdhury Cindy K. Bahler Cheryl R. Hann Bradley H. Holman Michael P. Fautsch
author_sort	Uttio Roy Chowdhury
title	Isolation and characterization of novel primary cells from the human distal outflow pathway
title_short	Isolation and characterization of novel primary cells from the human distal outflow pathway
title_full	Isolation and characterization of novel primary cells from the human distal outflow pathway
title_fullStr	Isolation and characterization of novel primary cells from the human distal outflow pathway
title_full_unstemmed	Isolation and characterization of novel primary cells from the human distal outflow pathway
title_sort	isolation and characterization of novel primary cells from the human distal outflow pathway
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/1cf5c2287ff2434d90daf7456fc0bb71
work_keys_str_mv	AT uttioroychowdhury isolationandcharacterizationofnovelprimarycellsfromthehumandistaloutflowpathway AT cindykbahler isolationandcharacterizationofnovelprimarycellsfromthehumandistaloutflowpathway AT cherylrhann isolationandcharacterizationofnovelprimarycellsfromthehumandistaloutflowpathway AT bradleyhholman isolationandcharacterizationofnovelprimarycellsfromthehumandistaloutflowpathway AT michaelpfautsch isolationandcharacterizationofnovelprimarycellsfromthehumandistaloutflowpathway
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Isolation and characterization of novel primary cells from the human distal outflow pathway

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