Porcine small intestinal submucosa (SIS): a bioscaffold supporting in vitro primary human epidermal cell differentiation and synthesis of basement membrane proteins.
The growth pattern of human epidermal cells, fibroblasts or Swiss mouse 3T3/J2 fibroblasts cultured upon the extracellular matrix (ECM) derived from small intestinal submucosa (SIS) was evaluated. The cell/SIS composites were grown submerged, then maintained in air/liquid interface for 2, 7, 10 or 14 days. The presence of differentiation-related keratins 10, 14 and 16, FN, laminin, collagen type VII and collagen type IV was determined by immunohistochemical methods in SIS alone and in the SIS/cell composite. Only FN could be detected in SIS alone. SIS supported the formation of an epithelial structure with suprabasal expression of K16 and regional suprabasal expression of K10. The epidermal cells were K14 positive and tended to 'invade' the SIS to various degrees. Following the growth of epidermal cells and fibroblasts on the SIS substratum, immunolabeling of FN, laminin, collagen type VII and collagen type IV was observed in a cell-associated pattern. The fibroblasts commonly invaded the SIS, when co-cultivated with epidermal cells on the opposite side of the SIS. The ability of SIS to support epidermal cell/fibroblast attachment, migration and/or proliferation and differentiation with deposition of basement membrane (BM) components indicates that the composite model may be useful for studying cell-matrix interactions and for investigation as a dermal substitute.
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Department of Biomedical Engineering, 1296 A.A. Potter Building, Room 204, Purdue University, West Lafayette, IN 47907-1296, USA. badylak@ecn.purdue.edu
Badylak
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D016475:3T3 Cells / Q000166:cytology / Q000378:metabolism; D000328:Adult; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000032:analysis; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000166:cytology*; D005353:Fibronectins / Q000032:analysis; D005612:Freeze Drying; D006801:Humans; D007150:Immunohistochemistry; D007231:Infant, Newborn; D007413:Intestinal Mucosa; D007421:Intestine, Small; D007633:Keratins / Q000032:analysis; D007797:Laminin / Q000032:analysis; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis*; D051379:Mice; D013552:Swine
D005353:Fibronectins; D007797:Laminin; D008565:Membrane Proteins; D007633:Keratins; D003094:Collagen