Cellular artificial skin substitute produced by short period simultaneous culture of fibroblasts and keratinocytes

Abstract

We developed a novel artificial skin substitute consisting of two collagen sponge layers with different pore sizes and cross-link densities. Fibroblasts suspended in 0.5 ml Dulbecco-modified Eagle's medium (DMEM) + 10% fetal bovine serum (FBS) were seeded on the lower dermal sponge layer, then epidermal collagen sponge and 0.1 ml suspension of keratinocytes in KGM were layered in this order. After a few hours, the medium was changed to DMEM + 5% FBS. These processes were carried out in one day, and the composite layers were then cultured by the air-liquid interface culture method. Three to five days after seeding, keratinocytes had grown to about ten layers, and fibroblasts had grown three-dimensionally into the lower dermal sponge layer. This novel cellular artificial skin substitute was grafted onto nude mice and took in 4 weeks. This skin substitute has the advantage of a shorter culturing period than previously cultured skins, and may be clinically useful for grafting that is urgently required in patients with severe generalised burns.

Allogeneic Transplantation of an Adipose-Derived Stem Cell Sheet Combined With Artificial Skin Accelerates Wound Healing in a Rat Wound Model of Type 2 Diabetes and Obesity

“…rASC sheets were successfully created from Lewis rats exhibiting a wide range of ages (8,10,16,20, and 33 weeks) (data not shown), and rASC sheets from 20-33-week-old rats were used for the transplantations. This study showed that rASC sheets from 33-week-old rats secreted growth factors and accelerated wound healing compared with the control group, suggesting that adipose tissue from middle-aged rats created functional rASC sheets.…”

“…Cell-based therapy has emerged as a new application for the treatment of ulcers (8,9). In particular, mesenchymal stem cells (MSCs) exhibit an excellent potential for increasing the rate of wound healing given their selfrenewal capacity, immunomodulatory effects, and ability to differentiate into various cell lineages (10).…”

Identification of cartilage progenitor cells in the adult ear perichondrium: utilization for cartilage reconstruction

“…21 The thickness of the sheet was adjusted to 2 mm, and the average pore size was 90 mm. Collagen sponge was cut into round disks with 12 mm diameter.…”

Immunoarchitectural characterization of a human skin model reconstructed in vitro

“…In a second stage, bovine collagen would be replaced by collagen from the injected fibroblasts, as accepted in other studies. 1,4 The presence of only a few elastic fibers in the skin model, compared with normal skin (Weigert-Van Gieson staining), was probably due to the short time for which the cells remained in the culture medium, as mentioned earlier. 15 The presence of a thin basement membrane in HSERIV (as seen via the PAS technique) indirectly demonstrates the formation of a completely differentiated skin equivalent.…”

“…[1][2][3][4][5][6] These models represent an alternative system for testing pharmacological products in the place of laboratory animals. 7 The European Union, through its "Sixth Amendment to the European Community Cosmetic Directive", has been aiming towards banning the use of animals in tests for certain types of cosmetic ingredients, combinations of ingredients and final products since January 1, 1998.…”