Epithelial tissue formation in matrix culture and in histophysiologic gradient culture

Leighton, Joseph

doi:10.1007/bf01409012

Cited by 2 publications

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“…Sterile Gelfoam sponges from purified pork skin gelatin (12 × 7 mm, product code NDC 0009-0315-03; Pharmacia & Upjohn; Kalamazoo, MI; http://www.pnu.com) were moisturized with KGM containing 10% FBS and KGS just before use and then transferred to six-well culture plates, on top of a monolayer of metabolically active, irradiated HPI.1 cells as feeders. Keratinocytes were placed on the top surface of each sponge (five injections, each of approximately 1 × l0 4 cells per 20 µl), as described elsewhere [44]. The cultures were then incubated in a stationary horizontal position in the incubator at 37°C in a 5% CO 2 -in-air atmosphere.…”

Section: Cultures On Gelatin Sponge Matricessupporting

confidence: 92%

Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long‐Term Culture

et al. 2003

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We developed a procedure for growing normal epidermal keratinocyte stem cells isolated from a single punch biopsy of adult human skin in long-term culture. Primary skin epithelial cells were maintained in collagen-coated plates with irradiated human neonatal foreskin fibroblasts (line HPI.1) as a feeder for more than 120 days, approximately 115 population doublings, without signs of replicative senescence. Clonal analysis revealed the presence of holoclones, meroclones, and paraclones. Only emerging colonies with high proliferative potentials and extensive capacities for division (holoclones and meroclones) were subcultured, favoring the expansion of stem cells and progenitors capable of prolonged self-maintenance when subcloned, thus accounting for the prevailing long-term proliferation of the original culture. We found that meroclones included bipotent progenitors capable of generating both keratinocytes and mucin-producing cells. The numbers of these cells were greater after confluence, suggesting that commitment for their differentiation occurred late in the life of a single clone. On a three-dimensional gelatin matrix and on a collagen layer containing the fibroblast feeder, cells isolated from the expansion of holoclones and meroclones formed stratified cohesive layers of keratinocytes that were able to further differentiate, as in normal skin. These results indicate that our procedure will serve as a valuable tool to study expansion of epidermal stem cells as well as the growth mechanisms and cell products associated with their growth and differentiation.

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Section: Cultures On Gelatin Sponge Matricessupporting

confidence: 92%

Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long‐Term Culture

et al. 2003

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Exploring processes of organization of normal and neoplastic epithelial tissues in gradient culture

Leighton¹

1994

J of Cellular Biochemistry

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The biology of animal cells in culture is often studied in individual cells or in sheets of cells. The relevance of such studies to the intact animal is unclear, since the spatial conditions encountered by cells in animals is one of dense three-dimensional masses of cells, with limits to migration, and with gradients both of diffusion of metabolites and of morphologic maturation. These spatial requisites have gradually been met in culture. A brief account describes sponge matrix culture for three-dimensional growth and unilaminar, bilaminar, and radial histophysiologic gradient cultures. Some of the common neoplastic abnormalities of surface epithelial tissues are considered. Proposals for investigating the histokinetic mechanisms regulating some epithelial tissue processes are suggested. In the most recent development of gradient culture methods, a thin permeable transparent collagen membrane is intrinsically strengthened by producing a waffle membrane pattern for histophysiologic gradient culture.

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Epithelial tissue formation in matrix culture and in histophysiologic gradient culture

Cited by 2 publications

References 12 publications

Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long‐Term Culture

Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long‐Term Culture

Exploring processes of organization of normal and neoplastic epithelial tissues in gradient culture

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