Abstract A method, solid-state culture support and culture solution are disclosed which enables successful in vitro culturing of differentiated cells, with significant retention of their differentiated character. Through the use of extracellular matrix fibers, specifically derived from connective tissue, as culture substrates, the method also discloses the isolation of the connective tissue fibers and their preparation as a culture substrate. This method provides significantly higher survival and attachment rates, and often significantly improved growth properties for in vitro cultures of differentiated cells, especially epithelial, over the current methods for culturing these cells. This method also significantly enables certain differentiated cells to retain much of their normal enzymatic activities. Furthermore, this method enables certain differentiated cells to retain to a high degree, their ability to secrete substances, such as hormones.
A cell culture environment consisting essentially of a base support for containing culture medium and said cells cultured thereon and a plurality of connective tissue-derived fibers in suspension within a solution conducive to culturing cells and within said base support, said connective tissue-derived fibers consisting essentially of collagens, non-collagenous proteins and carbohydrates.
Claim 2
Dependent Claim
52 words
A cell culture environment consisting essentially of a base support for containing culture medium and said cells cultured thereon and a plurality of connective tissue-derived fibers in suspension within a solution conducive to culturing cells and within said base support, said connective tissue-derived fibers consisting essentially of collagens, non-collagenous proteins and carbohydrates.
Claim 3
Dependent Claim
26 words
The cell culture environment of claim 1, further comprising a medium supplemented with hormones and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 4
Dependent Claim
26 words
The cell culture environment of claim 1, further comprising a medium supplemented with plasma and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 5
Dependent Claim
26 words
The cell culture environment of claim 1, further comprising a medium supplemented with serum and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 6
Dependent Claim
28 words
The cell culture environment of claim 1, further comprising a medium supplemented with hormones and plasma and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 7
Dependent Claim
28 words
The cell culture environment of claim 1, further comprising a medium supplemented with hormones and serum and with conditioned medium including factors derived from cultures of mesenchymal cells.
Claim 8
Dependent Claim
17 words
The cell culture environment of claim 1, wherein said connective tissue-derived fibers are derived from organ tissue.
Claim 9
Dependent Claim
25 words
The cell culture environment of claim 1, wherein said cells cultered in said environment attach to said connective tissue-derived fibers in suspension within the solution.