The University of North Carolina at Chapel Hill;Roach, Marsha Lynn;Malavarca, Richard Harold
发明人:
ROACH, Marsha Lynn,MALAVARCA, Richard Harold,WANG, Yunfang,REID, Lola Cynthia McAdams
申请号：
ES11801504
公开号：
ES2626145T3
申请日:
2011.07.01
申请国别(地区):
ES
年份:
2017
代理人:
摘要：
A method for producing a biomatrix scaffold from mammalian liver tissue for dispersion on an industrial scale on a culture apparatus； The method comprises: a) perfusing the liver tissue or homogenizing the liver tissue with a buffer comprising a salt concentration of 3.5 M NaCl to 4.5 M NaCl, then b) perfusing the liver tissue or extracting the homogenate of step (a) with a slip buffer comprising sodium deoxycholate and phospholipase A2 in a first medium, wherein the osmolality of said first medium is 250 mOsm / kg to 350 mOsm / kg and said first medium is serum free and at neutral pH； then c) perfuse the tissue or extract the homogenate from step (b) with a buffer at a neutral pH and comprising a salt concentration of 2.0 M NaCl to 5.0 M NaCl, the concentration chosen to maintain insoluble collagen identified in biological tissue； then d) perfuse the tissue or extract the homogenate from step (c) with RNAase and DNAase in a buffer； and then e) rinse the tissue or homogenate of step (d) with a second medium that is at neutral pH, is serum free and has an osmolality of 250 mOsm / kg to 350 mOsm / kg, to thereby produce a intact or homogenized biomatrix scaffold of liver tissue, said biomatrix scaffold retains at least 95% of its original collagen and most of the matrix components associated with collagen and growth factors, hormones and cytokines attached to the tissue matrix biological； f) dilute the biomatrix scaffold in basal medium； g) freeze the biomatrix scaffold of (f)； h) pulverize the biomatrix scaffold of (g) by cryogenic crushing until obtaining biomatrix particles having a size in the range of 1 μm to 100 μm； i) defrosting the biomatrix particles of (h) in suspension in basal medium； and j) dispersing the biomatrix particles of step (i) on a culture apparatus, to thereby produce a biomatrix scaffold from biological tissue for dispersion on an industrial scale on a culture apparatus.Un método para producir un andamio de biomatriz a partir de tejido hepátic
