Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor tissue

Jeffrey M.A. van der Krogt; Marchien G. Dallinga; Antoon J. van den Bogaerdt; Ivanka J.E. van der Meulen; Jaap D. van Buul

doi:10.1016/j.xpro.2026.104535

Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor tissue

Jeffrey M.A. van der Krogt^*
, Marchien G. Dallinga
, Antoon J. van den Bogaerdt
, Ivanka J.E. van der Meulen
, Jaap D. van Buul^*

^*Corresponding author for this work

Department of Ophthalmology

Research output: Contribution to journal › Article › Academic › peer-review

2 Downloads (Pure)

Abstract

Studying the function of healthy corneal endothelial cells has been challenging due to a variety of culturing strategies. Here, we present a standardized protocol for culturing primary human corneal endothelial cells, isolated from research-grade corneoscleral donor tissue, without the addition of mitotic enhancers. We describe the required enzymes, medium composition, cell density, and incubation times. Additionally, we use immunofluorescence microscopy to illustrate optimal outcomes and discuss troubleshooting procedures.

Original language	English
Article number	104535
Journal	STAR protocols
Volume	7
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2026.104535
Publication status	Published - 19 Jun 2026

Access to Document

10.1016/j.xpro.2026.104535Licence: CC BY

1-s2.0-S2666166726001887-mainFinal published version, 6.1 MBLicence: CC BY

Cite this

@article{1440a11408b6451b9d27fb1b16165ab4,

title = "Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor tissue",

abstract = "Studying the function of healthy corneal endothelial cells has been challenging due to a variety of culturing strategies. Here, we present a standardized protocol for culturing primary human corneal endothelial cells, isolated from research-grade corneoscleral donor tissue, without the addition of mitotic enhancers. We describe the required enzymes, medium composition, cell density, and incubation times. Additionally, we use immunofluorescence microscopy to illustrate optimal outcomes and discuss troubleshooting procedures.",

author = "\{van der Krogt\}, \{Jeffrey M.A.\} and Dallinga, \{Marchien G.\} and \{van den Bogaerdt\}, \{Antoon J.\} and \{van der Meulen\}, \{Ivanka J.E.\} and \{van Buul\}, \{Jaap D.\}",

note = "Publisher Copyright: {\textcopyright} 2026 The Authors.",

year = "2026",

month = jun,

day = "19",

doi = "10.1016/j.xpro.2026.104535",

language = "English",

volume = "7",

journal = "STAR protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor tissue

AU - van der Krogt, Jeffrey M.A.

AU - Dallinga, Marchien G.

AU - van den Bogaerdt, Antoon J.

AU - van der Meulen, Ivanka J.E.

AU - van Buul, Jaap D.

PY - 2026/6/19

Y1 - 2026/6/19

N2 - Studying the function of healthy corneal endothelial cells has been challenging due to a variety of culturing strategies. Here, we present a standardized protocol for culturing primary human corneal endothelial cells, isolated from research-grade corneoscleral donor tissue, without the addition of mitotic enhancers. We describe the required enzymes, medium composition, cell density, and incubation times. Additionally, we use immunofluorescence microscopy to illustrate optimal outcomes and discuss troubleshooting procedures.

AB - Studying the function of healthy corneal endothelial cells has been challenging due to a variety of culturing strategies. Here, we present a standardized protocol for culturing primary human corneal endothelial cells, isolated from research-grade corneoscleral donor tissue, without the addition of mitotic enhancers. We describe the required enzymes, medium composition, cell density, and incubation times. Additionally, we use immunofluorescence microscopy to illustrate optimal outcomes and discuss troubleshooting procedures.

UR - https://www.scopus.com/pages/publications/105037559639

U2 - 10.1016/j.xpro.2026.104535

DO - 10.1016/j.xpro.2026.104535

M3 - Article

AN - SCOPUS:105037559639

SN - 2666-1667

VL - 7

JO - STAR protocols

JF - STAR protocols

IS - 2

M1 - 104535

ER -

Protocol for culturing healthy primary endothelial cells isolated from research-grade human corneoscleral donor tissue

Abstract

Access to Document

Other files and links

Fingerprint

Cite this