×

Biomatrix scaffolds for industrial scale dispersal

  • US 9,102,913 B2
  • Filed: 07/01/2011
  • Issued: 08/11/2015
  • Est. Priority Date: 07/02/2010
  • Status: Active Grant
First Claim
Patent Images

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 μ

    m;

    (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.

View all claims
  • 4 Assignments
Timeline View
Assignment View
    ×
    ×