US 11,913,027 B2
Methods of treatment using pluripotent human adipose adult stem cells
Gregorio Chazenbalk, Studio City, CA (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA (US)
Filed by THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA (US)
Filed on Jul. 8, 2021, as Appl. No. 17/305,483.
Application 16/179,798 is a division of application No. 14/893,014, granted, now 10,131,880, issued on Nov. 20, 2018, previously published as PCT/US2014/039137, filed on May 22, 2014.
Application 17/305,483 is a continuation of application No. 16/179,798, filed on Nov. 2, 2018, granted, now 11,066,647.
Claims priority of provisional application 61/826,417, filed on May 22, 2013.
Prior Publication US 2021/0403872 A1, Dec. 30, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0775 (2010.01); A61K 35/28 (2015.01); C12N 5/074 (2010.01)
CPC C12N 5/0667 (2013.01) [A61K 35/28 (2013.01); C12N 5/0607 (2013.01); C12N 2500/02 (2013.01); C12N 2501/734 (2013.01); C12N 2506/1384 (2013.01)] 19 Claims
 
1. A method of treating traumatic injury or disease-associated damage in a subject, the method, comprising:
(i) isolating pluripotent stem cells (PASCs) by a method comprising:
(a) releasing adipose cells, including an adipocyte fraction and a stromal vascular fraction, from an adipose tissue sample using a proteolytic enzyme that breaks the peptide bonds in collagen;
(b) co-incubating the released adipocytes and the released stromal vascular fraction for 2-36 hours, wherein 4-24 hours of said co-incubation takes place under stress conditions in a medium containing a proteolytic enzyme, in the absence of nutrients, under hypoxic conditions, and decreasing the temperature to 4° C.;
(c) recovering viable cells following the co-incubation in step (b) by centrifuging to produce a cell pellet, removing supernatant containing adipocyte cell debris by aspiration, washing the cell pellet to remove the proteolytic enzyme, and resuspending the recovered cells in media wherein the recovered cells comprise PASCs; and
(ii) administering a composition comprising the PASCs to the subject under conditions permitting the PASCs of the composition to divide and populate a site of tissue damage.