文档介绍：Methods in Molecular Biology TM
VOLUME 188
EpithelialEpithelial
CellCell CultureCulture
ProtocolsProtocols
EditedEdited byby
ClareClare WiseWise
HUMANA PRESS
HLE Cell Culture 1
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Human Lens Epithelial Cell Culture
Nobuhiro Ibaraki
1. Introduction
Crystalline lens consists of epithelial cells, fiber cells, and a capsule. They
originate from lens epithelial cells. Epithelial cells differentiate into fiber cells
and produce collagen, which is the pound of the capsule. The lens
epithelial cells maintain normal physiology and homeostasis of the lens, so the
cultures of human lens epithelial (HLE) cells provide important information
concerning the role of epithelium in normal and cataract formation.
The difficulty of HLE cell culture is due to limited sources of the cells and a
low viability and delicacy of the cells. HLE cells are easily damaged, resulting
in the failure of the culture by mechanical injury, contamination, toxicity of
reagents, and freezing for storage.
This chapter describes the procedures of HLE cell culture, so that scientists
who are unfamiliar with this culture may carry it out essfully. The sources
of HLE cells, explant culture, harvest, subculture, storage, and shipment are
explained. The critical points for HLE cell culture are also stated in the note
section.
2. Materials
1. o’s modified Eagle medium (DMEM).
2. Fetal bovine serum (FBS), qualified (see Note 2).
3. Gentamicin reagent solution.
4. Growth medium: DMEM supplemented with FBS is used as a standard medium.
As the viability of HLE cells is very low (see Note 3), neither antibiotic nor
antifungal agents should be used for HLE cell culture except cell line cell culture.
Gentamicin reagent solution (10 µg/mL) can be used for HLE cell line cell
From: Methods in Molecular Biology, vol. 188: Epithelial Cell Culture Protocols
Edited by: C. Wise © Humana Press Inc., Totowa, NJ
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2Ibaraki
culture. The concentration of serum should be at