Effect of Physiological Oxygen on Primary Human Corneal Endothelial Cell Cultures.
Effect of Physiological Oxygen on Primary Human Corneal Endothelial Cell Cultures.
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DOI：
10.1167/tvst.11.2.33
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发表时间：
2022-02-01
影响因子：
3
通讯作者：
Shi MY
中科院分区：
文献类型：
--
作者：
Patel SP;Calle Gonzalez B;Paone N;Mueller C;Floss JC;Sousa ME;Shi MY
Primary human corneal endothelial cells (HCEnCs) cultured in room air are exposed to significantly higher O2 concentrations [O2] than what is normally present in the eye. We evaluated the growth and metabolism of HCEnCs cultured under physiological [O2] (2.5%; [O2]2.5) and room air ([O2]A). Primary cultures of HCEnCs from normal donors and donors with Fuchs dystrophy were grown at [O2]2.5 and [O2]A. Growth and morphology were compared using phase-contrast microscopy, zonula occludens (ZO-1) localization, cell density measurements, and senescence marker staining. CD44 (cell quality) and HIF-1α (hypoxia-inducible factor-1α) levels were evaluated by Western blotting. Cell adaptability to a reversal of [O2] growth conditions was measured with cell viability assays, and cell metabolism was assessed via oxygen consumption and extracellular acidification rates. HCEnCs grown at [O2]A and [O2]2.5 displayed similar morphologies, ZO-1 localization, CD44 expression, and senescence. Cells from donors with Fuchs dystrophy grew better at [O2]2.5 than at [O2]A. HIF-1α was undetectable. Cells displayed greater viability at [O2]2.5 than at [O2]A. HCEnCs showed significantly greater proton leak (P < 0.01), nonmitochondrial oxygen consumption (P < 0.01), and spare capacity (P < 0.05) for oxygen consumption rates, and greater basal glycolysis (P < 0.05) with a decreased glycolytic reserve capacity (P < 0.05) for extracellular acidification rates. Primary HCEnCs show unique metabolic characteristics at physiologic [O2]. The effect of [O2] for optimization of HCEnC culture conditions should be considered. With the advance of cell-based therapeutics for corneal endothelial diseases, [O2] should be considered an important variable in the optimization of HCEnC culture conditions.
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影响因子：
7.8
作者：
Lowe D;Raj K
通讯作者：
Raj K
影响因子：
4.4
作者：
Joyce, Nancy C.;Harris, Deshea L.;Zhu, Cheng C.
通讯作者：
Zhu, Cheng C.
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3.5
作者：
Braunschweig, Lena;Meyer, Anne K.;Storch, Alexander
通讯作者：
Storch, Alexander
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3.4
作者：
Matthaei, Mario;Zhu, Angela Y.;Kallay, Laura;Eberhart, Charles G.;Cursiefen, Claus;Jun, Albert S.
通讯作者：
Jun, Albert S.
影响因子：
4.8
作者：
Baillet, Athan;Hograindleur, Marc-Andre;Paclet, Marie-Helene
通讯作者：
Paclet, Marie-Helene