The Cultivation of Lens Epithelium in Vitro

Kirby, Daniel B.

doi:10.1084/jem.45.6.1009

Cited by 17 publications

(4 citation statements)

References 7 publications

(8 reference statements)

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“…LECs were first cultured from embryonic chicken by Kirby in 1927 3 . Since then, chicken LECs cultures have been widely used and significantly improved upon, especially by Menko et al .…”

Section: Introductionmentioning

confidence: 99%

Roles of TGFβ and FGF signals during growth and differentiation of mouse lens epithelial cell in vitro

Wang

Liu

et al. 2017

Sci Rep

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Transforming growth factor β (TGFβ) and fibroblast growth factor (FGF) signaling pathways play important roles in the proliferation and differentiation of lens epithelial cells (LECs) during development. Low dosage bFGF promotes cell proliferation while high dosage induces differentiation. TGFβ signaling regulates LEC proliferation and differentiation as well, but also promotes epithelial-mesenchymal transitions that lead to cataracts. Thus far, it has been difficult to recapitulate the features of germinative LECs in vitro. Here, we have established a LEC culture protocol that uses SB431542 (SB) compound to inhibit TGFβ/Smad activation, and found that SB treatment promoted mouse LEC proliferation, maintained LECs’ morphology and distinct markers including N-cadherin, c-Maf, Prox1, and αA-, αB-, and β-crystallins. In contrast, low-dosage bFGF was unable to sustain those markers and, combined with SB, altered LECs’ morphology and β-crystallin expression. We further found that Matrigel substrate coatings greatly increased cell proliferation and uniquely affected β-crystallin expression. Cultured LECs retained the ability to differentiate into γ-crystallin-positive lentoids by high-dosage bFGF treatment. Thus, a suppression of TGFβ/Smad signaling in vitro is critical to maintaining characteristic features of mouse LECs, especially expression of the key transcription factors c-Maf and Prox1.

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“…LECs were first cultured from embryonic chicken by Kirby in 1927 3 . Since then, chicken LECs cultures have been widely used and significantly improved upon, especially by Menko et al .…”

Section: Introductionmentioning

confidence: 99%

Roles of TGFβ and FGF signals during growth and differentiation of mouse lens epithelial cell in vitro

Wang

Liu

et al. 2017

Sci Rep

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“…Specialized techniques are required to study the limited population of lens epithelium monolayer cells. Lens epithelial cell differentiation and behavior have been studied in many species using in vitro primary cell culture and immortalized cell lines ( Kirby, 1927 ; Mann, 1948 ; Mamo and Leinfelder, 1958 ; Van Der Veen and Heyen, 1959 ; Okada et al, 1971 ; Creighton et al, 1976 ; Hamada and Okada, 1978 ; Reddan et al, 1980 ; Menko et al, 1984 ; FitzGerald and Goodenough, 1986 ; Reddan et al, 1989 ; Musil et al, 1990 ; Bermbach et al, 1991 ; Andley et al, 1998 ; Ibaraki et al, 1998 ; Ogiso et al, 1998 ; Wang et al, 2007 ; Bannik et al, 2013 ; Matsuyama et al, 2013 ; Sundelin et al, 2014 ; Terrell et al, 2015 ; Wang et al, 2017 ; Weatherbee et al, 2019 ). However, primary lens epithelial cells in culture often do not maintain native lens epithelial cell characteristics.…”

Section: Introductionmentioning

confidence: 99%

Methodologies to unlock the molecular expression and cellular structure of ocular lens epithelial cells

Parreno

Emin

et al. 2022

Front. Cell Dev. Biol.

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The transparent ocular lens in the anterior chamber of the eye is responsible for fine focusing of light onto the retina. The lens is entirely cellular with bulk of the tissue composed of fiber cells, and the anterior hemisphere of the lens is covered by a monolayer of epithelial cells. Lens epithelial cells are important for maintaining fiber cell homeostasis and for continual growth of the lens tissue throughout life. Cataracts, defined as any opacity in the lens, remain the leading cause of blindness in the world. Following cataract surgery, lens epithelial cells can undergo a process of epithelial-to-mesenchymal transition (EMT), leading to secondary cataracts due to posterior capsular opacification (PCO). Since the epithelial cells make up only a small fraction of the lens, specialized techniques are required to study lens epithelial cell biology and pathology. Studies using native lens epithelial cells often require pooling of samples to obtain enough cells to make sufficient samples for traditional molecular biology techniques. Here, we provide detailed protocols that enable the study of native mouse lens epithelial cells, including immunostaining of the native lens epithelium in flat mounts, extraction of RNA and proteins from pairs of lens epithelial monolayers, and isolation of lens epithelial cells for primary culture. These protocols will enable researchers to gain better insight on representative molecular expression and cellular structure of lens epithelial cells. We also provide comparative data between native, primary culture, and immortalized lens epithelial cells and discuss the advantages and disadvantages of each technique presented.

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“…Schon seit mehr als 70 Jahren werden Epithelzellen der Augenlinse kultiviert. Kirby [26] hat bei seinen Kultivierungsversuchen Linsen von Hühnerembryonen verwendet und fand, dass das unter der vorderen Linsenkapsel lokalisierte Epithel zu mitotischem Wachstum fähig ist. In den folgenden Jahren wurden Linsenepithelzellen von verschiedenen Tierspezies (Rind, Kaninchen, Ratte, Primaten) in Kultur genommen.…”

unclassified

Humane Linsenzellen in der Kultur12 - I. Gewinnung von adulten Linsenepithelzellen aus Linsenkapselpräparaten nach Kataraktoperationen und von fetalen Linsenzellen durch Reaktivierung kernhaltiger Faserzellen -

Iwig¹,

Glässer²,

Luther³

et al. 2001

Klin Monatsbl Augenheilkd

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Lens-capsule-preparations available from cataract surgery are well suited for the isolation of human lens cell lines, which were needed for testing cytotoxicity of drugs and for tracing of cataractogenic risk factors. The finding that nucleus containing fiber cells from the equator of human lenses can be reactivated to proliferating cells let us suppose, that these cells, which can not be removed easily from the posterior lens capsule, contribute to the after cataract formation.

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The Cultivation of Lens Epithelium in Vitro

Cited by 17 publications

References 7 publications

Roles of TGFβ and FGF signals during growth and differentiation of mouse lens epithelial cell in vitro

Roles of TGFβ and FGF signals during growth and differentiation of mouse lens epithelial cell in vitro

Methodologies to unlock the molecular expression and cellular structure of ocular lens epithelial cells

Humane Linsenzellen in der Kultur12 - I. Gewinnung von adulten Linsenepithelzellen aus Linsenkapselpräparaten nach Kataraktoperationen und von fetalen Linsenzellen durch Reaktivierung kernhaltiger Faserzellen -

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