Patent ID: 9102913
Filing Date: 2015-08-11
Classification: A61L,C12N

Abstract:
1. A method of producing a biomatrix scaffold from biological tissue for industrial scale dispersal onto culture apparatus, comprising: (a) perfusing or homogenizing biological tissue with a buffer comprising a salt concentration from about 3.5 M NaCl to about 4.5 M NaCl, wherein the tissue is selected from the group consisting of liver, lung, thyroid, skin, pancreas, blood vessels, bladder, kidneys, brain, biliary tree, duodenum, abdominal aorta, iliac vein, heart and intestines; then (b) perfusing the biological tissue or extracting the homogenate of step (a) with a delipidating buffer comprising lipases and detergents in a serum free first medium, having a neutral pH and an osmolality from about 250 mOsm/kg to about 350 mOsm/kg, wherein the detergents do not comprise sodium dodecyl sulfate or TritonX-100; then (c) perfusing the tissue or extracting the homogenate of step (b) with a buffer at a neutral pH and comprising a salt concentration from about 2.0 M NaCl to about 5.0 M NaCl, the concentration chosen to keep insoluble collagens identified in the biological tissue; then (d) perfusing the tissue or extracting the homogenate of step (c) with RNase and DNase in a buffer; and then (e) rinsing the tissue or homogenate of step (d) with a second medium that is at neutral pH, is serum-free and has an osmolality from about 250 mOsm/kg to about 350 mOsm/kg, thereby producing an intact or homogenized biomatrix scaffold from the biological tissue, said biomatrix scaffold comprising at least 95% of the collagens and most collagen-associated matrix components and matrix bound growth factors, hormones and cytokines of the biological tissue; (f) diluting the biomatrix scaffold in basal medium; (g) freezing the biomatrix scaffold of (f); (h) pulverizing the biomatrix scaffold of (g) by cryogenic grinding into biomatrix particles ranging in size from about 1 nm to about 100 μm; (i) thawing the biomatrix particles of (h) in suspension in basal medium; and (j) dispersing the biomatrix particles of step (i) onto a culture apparatus, thereby producing a biomatrix scaffold from biological tissue for industrial scale dispersal onto culture apparatus.