US 12,343,447 B2
Methods for tissue decellularization
Paola Bonfanti, London (GB); and Asllan Gjinovci, London (GB)
Assigned to UCL BUSINESS LTD, London (GB)
Appl. No. 17/055,328
Filed by UCL BUSINESS LTD, London (GB)
PCT Filed May 14, 2019, PCT No. PCT/GB2019/051310
§ 371(c)(1), (2) Date Nov. 13, 2020,
PCT Pub. No. WO2019/220091, PCT Pub. Date Nov. 21, 2019.
Claims priority of application No. 1807788 (GB), filed on May 14, 2018.
Prior Publication US 2021/0187164 A1, Jun. 24, 2021
Int. Cl. A61L 27/36 (2006.01); A61L 27/38 (2006.01); C12N 5/00 (2006.01); C12N 5/0775 (2010.01); C12N 5/078 (2010.01); G01N 33/50 (2006.01)
CPC A61L 27/3687 (2013.01) [A61L 27/3633 (2013.01); A61L 27/3813 (2013.01); A61L 27/3834 (2013.01); A61L 27/3886 (2013.01); C12N 5/0068 (2013.01); C12N 5/0634 (2013.01); C12N 5/0662 (2013.01); G01N 33/5088 (2013.01); C12N 2502/11 (2013.01); C12N 2502/13 (2013.01); C12N 2533/90 (2013.01)] 15 Claims
 
1. A method of producing a decellularised extracellular matrix (ECM) scaffold of at least a portion of a lobular organ with no common artery, the method comprising:
a) closing afferent blood vessels to substantially seal a target lobular organ, or a portion thereof, with no common artery within a non-human donor or a dead/brain dead human donor;
b) optionally: (i) cleaning coagulum and/or blood from at least a portion of the closed afferent blood vessels; and/or (ii) perfusing the target lobular organ or the portion thereof to confirm closure of the afferent blood vessels to form a sealed organ or sealed portion thereof;
c) removing the sealed organ or the sealed portion thereof from the non-human donor or the dead/brain dead human donor; and
d) perfusing the sealed organ or the sealed portion thereof with a detergent and enzymatic solutions to obtain the decellularised ECM scaffold.
 
13. A method for producing an artificial organ, the method comprising:
i) producing a decellularised extracellular matrix (ECM) scaffold of at least a portion of a lobular organ with no common artery, the method comprising:
a) closing afferent blood vessels to substantially seal a target lobular organ, or a portion thereof, with no common artery within a non-human donor or a dead/brain dead human donor;
b) optionally: (i) cleaning coagulum and/or blood from at least a portion of the closed afferent blood vessels; and/or (ii) perfusing the target lobular organ or the or sealed portion thereof to confirm closure of the afferent blood vessels to form a sealed organ;
c) removing the sealed organ or the sealed portion thereof from the non-human donor or the dead/brain dead human donor; and
d) perfusing the sealed organ or the sealed portion thereof with a detergent and enzymatic solutions to obtain the decellularised ECM scaffold;
ii) repopulating the decellularised ECM scaffold with stromal cells, to form a repopulated scaffold, and
iii) culturing the repopulated scaffold in vitro for about 4 to about 7 days, thereby producing the artificial organ.