Biomatrix scaffolds for industrial scale dispersal
First Claim
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1. A method of producing a plurality of biomatrix particles from a biological tissue for dispersal onto culture apparatus, comprising:
- (a) perfusing or homogenizing the biological tissue with a buffer comprising a salt concentration from about 3.5 M NaCl to about 4.5 M NaCl;
(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;
(c) perfusing the biological tissue or extracting the homogenate of step (b) with a buffer at a neutral pH and comprising a salt concentration ranging from about 2.0 M NaCl to about 5.0 M NaCl, the concentration chosen to not lose insoluble collagens identified in the biological tissue;
(d) perfusing the biological tissue or extracting the homogenate of step (c) with RNase and DNase in a buffer;
(e) rinsing the biological tissue or homogenate of step (d) with a second medium having a neutral pH, is serum-free and having an osomolality ranging from about 250 mOsm/kg to about 350 mOsm/kg, thereby producing an intact or homogenized biomatrix scaffold from the biological tissue;
(f) pulverizing the intact or homogenized biomatrix scaffold into a plurality of biomatrix particles ranging in size from about 1 nm to about 100 μ
m.
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Abstract
The present invention provides biomatrix scaffolds for industrial scale dispersal.
19 Citations
16 Claims
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1. A method of producing a plurality of biomatrix particles from a biological tissue for dispersal onto culture apparatus, comprising:
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(a) perfusing or homogenizing the biological tissue with a buffer comprising a salt concentration from about 3.5 M NaCl to about 4.5 M NaCl; (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; (c) perfusing the biological tissue or extracting the homogenate of step (b) with a buffer at a neutral pH and comprising a salt concentration ranging from about 2.0 M NaCl to about 5.0 M NaCl, the concentration chosen to not lose insoluble collagens identified in the biological tissue; (d) perfusing the biological tissue or extracting the homogenate of step (c) with RNase and DNase in a buffer; (e) rinsing the biological tissue or homogenate of step (d) with a second medium having a neutral pH, is serum-free and having an osomolality ranging from about 250 mOsm/kg to about 350 mOsm/kg, thereby producing an intact or homogenized biomatrix scaffold from the biological tissue; (f) pulverizing the intact or homogenized biomatrix scaffold into a plurality of biomatrix particles ranging in size from about 1 nm to about 100 μ
m. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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Specification