Chemistry: molecular biology and microbiology – Apparatus – Bioreactor
Patent
1997-03-25
2000-07-11
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Apparatus
Bioreactor
435 4, 4352884, 435 721, 435 723, 4352841, 4352871, 4352879, 4352971, 4352991, 4353041, 435391, C12M 300, C12M 304, C12Q 100, C12N 500
Patent
active
060871571
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to the culturing of eukaryotic cells. More particularly, this invention is directed to a method for supporting the growth and tissue differentiation of eukaryotic cells in vitro, by contacting the eukaryotic cells with submucosal tissue of a warm-blooded vertebrate, under conditions conducive to eukaryotic cell proliferation.
BACKGROUND AND SUMMARY OF THE INVENTION
Tissue culture allows the study in vitro of animal cell behavior in an investigator-controlled physiochemical environment. However, cellular morphology and metabolic activity of cultured cells are affected by the composition of the substrate on which they are grown. Presumably cultured cells function best (i.e. proliferate and perform their natural in vivo functions) when cultured on substrates that closely mimic their natural environment. Currently, studies in vitro of cellular function are limited by the availability of cell growth substrates that present the appropriate physiological environment for proliferation and development of the cultured cells.
The ability of complex substrates to support cell growth in vitro has been previously reported, and matrix products supporting such growth are commercially available. For example, Becton Dickinson currently offers two such products: Human Extracellular Matrix and MATRIGEL.RTM. Basement Membrane Matrix. Human Extracellular Matrix is a chromatographically partially purified matrix extract derived from human placenta and comprises laminin, collagen IV, and heparin sulfate proteoglycan. (Kleinman, HK et al., U.S. Pat. No. 4,829,000 (1989)) MATRIGEL.RTM. is a soluble basement membrane extract of the Engelbreth-Holm-Swarm (EHS) tumor, gelled to form a reconstituted basement membrane. Both of these matrix products require costly biochemical isolation, purification, and synthesis techniques and thus production costs are high.
The present invention is directed to the use of vertebrate submucosa-derived matrices as substrates for the growth and attachment of a wide variety of cell types. The collagenous matrices for use in accordance with the present invention comprise highly conserved collagens, glycoproteins, proteoglycans, and glycosaminoglycans in their natural configuration and natural concentration. The extracellular collagenous matrix for use in this invention is derived from submucosal tissue of a warm-blooded vertebrate. Submucosal tissue can be obtained from various sources, including intestinal tissue harvested from animals raised for meat production, including, for example, pigs, cattle and sheep or other warm-blooded vertebrates. This tissue can be used in either its natural configuration or in a comminuted or partially digested fluidized form. Vertebrate submucosal tissue is a plentiful by-product of commercial meat production operations and is thus a low cost cell growth substrate, especially when the submucosal tissue is used in its native layer sheet configuration.
The submucosal cell growth substrates of this invention provide cells with a collagenous matrix environment in vitro resembling that found in vivo. The natural composition and configuration of submucosal tissue provides a unique cell growth substrate that promotes the attachment and proliferation of cells.
Accordingly, one object of the present invention is to provide a relatively inexpensive cell culture growth substrate that promotes or induces growth and differentiation of cells cultured in vitro.
Another object of this invention is to provide a method for improving cell proliferation in cell/tissue culture by using vertebrate submucosal tissue as a substrate for cell/tissue growth in vitro.
Another object of this invention is to provide a cell culture composition including a proliferating cell population in contact with submucosal tissue of a warm-blooded vertebrate and a nutrient medium for support of the growth of said cell population.
Still a further object of this invention is to provide a model system for studying tumor cell growth. The model system comprises a p
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17-Estrodiol Inhibits IL-6 Production by Bone Marrow Stromal Cells and Osteoblasts In Vitro: A Potential
Badylak Stephen F.
Boder George
Critser John K.
Demeter Robert J.
Liu Chi
Clarian Health Partners
Lankford , Jr. Leon B.
Purdue Research Foundation
Tate Christopher R.
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