# US 9,752,115 B2 · Culture conditions for expansion of nephron progenitor cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,752,115 B2 - Document ID: US9752115B2 - Status: Granted - Kind: B2 - Application number: 14/632,454 - Filed: 2015-02-26 - Published: 2015-10-01 - Granted: 2017-09-05 - Earliest claimed: 2014-02-26 - Assignees: Maine Medical Center Research Institute - Applicants: Maine Medical Center Research Institute - Inventors: Leif Oxburgh; Aaron Brown - Web page: https://patentgrep.com/patent/US9752115B2 ## CPC - C12N 5/0018: {Culture media for cell or tissue culture (media for specific animal cell type C12N5/06)} - C12N 5/0686: {Kidney cells} - C12N 5/0687: {Renal stem cells; Renal progenitors} - C12N 2501/105: Insulin-like growth factors [IGF] - C12N 2501/119: Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10 - C12N 2501/155: Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor - C12N 2501/415: Wnt; Frizzeled - C12N 2501/727: Kinases (EC 2.7.) - C12N 2501/91: Heparin - C12N 2501/999: Small molecules not provided for elsewhere - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to compositions and methods associated with cocktails of growth factors and small molecules that target specific cell signaling pathways, the cocktails having been formulated to allow/promote the expansion of progenitor cells (e.g., nephron progenitor cells) within a defined culture system. ## Claims ### Claim 1 A composition for expanding a mammalian progenitor cell population comprising FGF9 at 25 to 250 ng/ml, LDN-193189 at 5 to 300 nM, CHIR 99021 at 100 to 10,000 nM and at least one of BMP7 and BMP4. ### Claim 2 (depends on 1) The composition of claim 1, wherein said mammalian progenitor cell population is a nephron progenitor cell population. ### Claim 3 (depends on 1) The composition of claim 1, further comprising ROCKi. ### Claim 4 (depends on 1) The composition of claim 1, further comprising at least one of IGF1, IGF2 and Heparin. ### Claim 5 (depends on 1) The composition of claim 1, further comprising one or more compositions selected from the group consisting of APEL or DMEM/F12+KOSR as media and matrigel or gelatin as ECM (extracellular matrix). ### Claim 6 (depends on 1) The composition of claim 1 comprising at least one component selected from the group consisting of: BMP7 (Bone morphogenetic protein 7, aka osteogenic protein-1 or OP-1) present at 5 to 100ng/ml; BMP4 (Bone morphogenetic protein 4) present at 5 to 100 ng/ml; ROCKi at 1 to 10 μM; IGF1 (Insulin-like growth factor 1, also called somatomedin C) present at 20 ng/ml; and IGF2 (Insulin-like growth factor 2) present at 2 ng/ml. ### Claim 7 (depends on 6) The composition of claim 6, comprising at least one component selected from the group consisting of: FGF9 (glia activating factor) present at 200 ng/ml; BMP7 (Bone morphogenetic protein 7, aka osteogenic protein-1 or OP-1) present at 30ng/ml; BMP4 (Bone morphogenetic protein 4) present at 30 ng/ml; LDN (BMP inhibitor LDN-193189) present at 75 nM; and CHIR 99021 present at 1 μM. ### Claim 8 (depends on 1) The composition of claim 1, comprising at least one component selected from the group consisting of IGF1 present at 5 to 100 ng/ml; IGF2 present at 0.1 to 10 ng/ml; and Heparin present at 1 to 10 μg/mg. ### Claim 9 (depends on 1) The composition of claim 1, comprising at least one component selected from the group consisting of IGF1 present at 20 ng/ml; IGF2 present at 2 ng/ml; Heparin present at 1 μg/mg; and ROCKi present at 10 μM. ### Claim 10 (depends on 1) A method for expanding a mammalian progenitor cell population comprising: a) obtaining a mammalian progenitor cell population; b) contacting said mammalian progenitor cell population with a composition of claim 1; and c) incubating said mammalian progenitor cell population with said composition for a time sufficient to allow for expansion of said mammalian progenitor cell population, thereby expanding said mammalian progenitor cell population. ### Claim 11 (depends on 10) The method of claim 10, wherein said mammalian progenitor cell population is a nephron progenitor cell population. ### Claim 12 (depends on 10) The method of claim 10, wherein said mammalian progenitor cell population is expanded by an amount selected from the group consisting of at least 10-fold, at least 100-fold, 256-fold, at least 1000-fold, 4096-fold and at least 5000-fold. ### Claim 13 (depends on 10) The method of claim 10, wherein said mammalian progenitor cell population is derived from embryonic stem cells. ### Claim 14 (depends on 10) The method of claim 10, wherein said mammalian progenitor cell population is human or murine. ### Claim 15 (depends on 10) The method of claim 10, wherein said incubating occurs for a duration of time selected from at least 18 hours, at least 2 days, at least 4 days and at least 9 days. ### Claim 16 (depends on 10) The method of claim 10, wherein said mammalian progenitor cell population has a doubling time of 18 hours. ### Claim 17 (depends on 10) The method of claim 10, wherein said cells are passaged every three days after four doublings. ### Claim 18 (depends on 1) A method for expanding a mammalian progenitor cell population within a mammalian stem cell population comprising: a) obtaining a mammalian stem cell population comprising mammalian progenitor cells; b) contacting said mammalian stem cell population with a composition of claim 1; and c) incubating said mammalian stem cell population with said composition for a time sufficient to allow for expansion of said mammalian progenitor cell population within said mammalian stem cell population, thereby expanding said mammalian progenitor cell population within said mammalian stem cell population. ### Claim 19 (depends on 18) The method of claim 18, wherein said mammalian progenitor cell population is a nephron progenitor cell population. ### Claim 20 (depends on 18) The method of claim 18, wherein said mammalian progenitor cell population is expanded by an amount selected from the group consisting of at least 2-fold, at least 5-fold, at least 10-fold and at least 20-fold. ### Claim 21 (depends on 18) The method of claim 18, wherein said mammalian progenitor cell population is initially present at less than 5% within said mammalian stem cell population. ### Claim 22 (depends on 18) The method of claim 18, wherein said mammalian progenitor cell population comprises at least 50% of all cells after incubating said mammalian stem cell population with said composition for a time sufficient to allow for expansion of said mammalian progenitor cell population within said mammalian stem cell population.