US 12,247,224 B2
Organoids for drug screening and personalized medicine
Ling Huang, Toronto (CA); and Senthil Muthuswamy, Toronto (CA)
Assigned to Senthil Muthuswamy, Toronto (CA); and Ling Huang, Toronto (CA)
Appl. No. 15/500,240
Filed by SENTING, LLC, Chestnut Hill, MA (US)
PCT Filed Jul. 30, 2015, PCT No. PCT/CA2015/050723
§ 371(c)(1), (2) Date Jan. 30, 2017,
PCT Pub. No. WO2016/015158, PCT Pub. Date Feb. 4, 2016.
Claims priority of provisional application 62/139,831, filed on Mar. 30, 2015.
Claims priority of provisional application 62/030,999, filed on Jul. 30, 2014.
Prior Publication US 2017/0267977 A1, Sep. 21, 2017
Int. Cl. C12N 5/071 (2010.01); C12N 5/00 (2006.01); C12N 5/09 (2010.01); G01N 33/50 (2006.01)
CPC C12N 5/0678 (2013.01) [C12N 5/0062 (2013.01); C12N 5/0676 (2013.01); C12N 5/0693 (2013.01); G01N 33/5011 (2013.01); G01N 33/507 (2013.01); G01N 33/5088 (2013.01); C12N 2500/38 (2013.01); C12N 2501/11 (2013.01); C12N 2501/115 (2013.01); C12N 2501/119 (2013.01); C12N 2501/15 (2013.01); C12N 2501/16 (2013.01); C12N 2501/33 (2013.01); C12N 2501/385 (2013.01); C12N 2501/999 (2013.01); C12N 2506/02 (2013.01); C12N 2533/90 (2013.01)] 18 Claims
 
1. A method for generating human pancreatic exocrine lineage organoids from pluripotent stem cell-derived pancreatic lineage committed progenitors, comprising:
a) digesting the pancreatic progenitors isolated from a sample;
b) plating the progenitors in a Pancreatic Progenitor and Tumor Organoid Medium (PTOM) comprising:
a cell culture medium selected from DMEM, F12, L-15, or RPMI, or combinations thereof;
an antioxidant selected from vitamin A or its derivatives, Resveratrol, Fisetin, or L-Glutathione;
a serum free supplement;
an insulin receptor agonist;
a glucocorticoid;
an FGFR agonist; and
one or more of an antibiotic, a retinoic receptor agonist, or an EGFR agonist;
c) replacing the PTOM with a Pancreatic Organoid Differentiation Media I (PODM I) comprising:
a cell culture medium selected from DMEM, F12, L-15, or RPMI, or combinations thereof;
an antioxidant;
a FGF;
an EGF;
a TGF-beta inhibitor; and
d) replacing the PODM I with a Pancreatic Organoid Differentiation Media II (PODM II) comprising:
a cell culture medium selected from DMEM, F12, L-15, or RPMI, or combinations thereof;
an antioxidant;
a FGF;
an EGF; and
a serum free supplement.