The matrix form of collagen and basal microporosity influence basal lamina deposition and laminin synthesis/secretion by stratified human keratinocytes in vitro
In vitro cellular & developmental biology. Animal(1995)
摘要
Summary The ability of the collagen matrix form to support the formation of a basal lamina by cultured normal human epidermal keratinocytes
(NHEK) was determined using transmission electron microscopy. The collagen matrix forms tested in this study were a) a dry
type I collagen film and b) a type I collagen gel. NHEK were grown for 14 days on the following five different substrates:
plain plastic culture dishes without the addition of collagen (PP); plain plastic culture dishes overlaid with a dry, aldehyde-crosslinked
type I collagen film (DCF-P); plain plastic culture dishes overlaid with an aldehyde-crosslinked type I collagen gel (GEL-P);
Millipore Millicell CM microporous membranes overlaid with a dry, aldehyde-crosslinked type I collagen film (DCF-CM); and
Millipore Millicell CM microporous membranes overlaid with an aldehyde-crosslinked type I collagen gel (GEL-CM). NHEK maintained
for 2 wk on PP and DCF-P were unable to secrete a basal lamina. NHEK grown for 2 wk on the GEL-P and GEL-CM substrates, however,
secreted a contiguous basal lamina at the GEL-NHEK interface. To determine if the appearance of this basal lamina correlated
with laminin synthesis, laminin was immunoprecipitated from cellular extracts, as well as media from the apical and basal
chambers. NHEK grown on the GEL-P substrate synthesized more laminin than did NHEK grown on the other four alternative substrates.
In addition, NHEK grown on GEL-CM were able to direct more laminin to the basal compartment than NHEK grown on DCF-CM substrates.
Taken together, the data indicate that the matrix form of collagen can influence basal lamina deposition, laminin synthesis,
and laminin trafficking in NHEK.
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关键词
microporous membrane.,keratinocytes,laminin,basal lamina,human epidermal models,transmission electron microscopy
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