# US 2017/0283777 A1 · Mammalian chimeric complementation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2017/0283777 A1 - Document ID: US20170283777A1 - Status: Published application - Kind: A1 - Application number: 15/468,006 - Filed: 2017-03-23 - Published: 2017-10-05 - Earliest claimed: 2016-03-25 - Assignees: The Salk Institute for Biological Studies - Applicants: Salk Institute for Biological Studies - Inventors: Juan Carlos IZPISUA BELMONTE; Jun WU - Web page: https://patentgrep.com/patent/US20170283777A1 ## CPC - C12N 5/0604: {Whole embryos; Culture medium therefor} - A01K 67/0271: Chimeric vertebrates, e.g. comprising exogenous cells - C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS} - A01K 2207/12: Animals modified by administration of exogenous cells - A01K 2227/101: Bovine - A01K 2227/108: Swine - C12N 2501/999: Small molecules not provided for elsewhere - C12N 2506/1307: from adult fibroblasts - C12N 2510/00: Genetically modified cells - C12N 2800/108: episomal vectors - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS ## Abstract Disclosed herein are chimeric mammals, such as a chimeric mammal comprising cells derived from at least a first mammal and a second mammal, wherein the cells from the first mammal comprise a genetic modification at one or more loci and the cells from the second mammal form at least one organ or tissue, wherein the first and second mammals are different species. ## Claims ### Claim at position 1 -23. (canceled) ### Claim 24 A mammalian organ or tissue isolated from a chimeric mammal comprising cells derived from at least a first mammal and a second mammal, wherein the organ or tissue is derived from an induced pluripotent stem cell (iPSC) cell from the second mammal, wherein the first and second mammal are different species. ### Claim 25 (depends on 24) The mammalian organ or tissue of claim 24, wherein the organ or tissue comprises at least one of the group consisting of liver, kidney, pancreas, hematopoietic stem cells, spleen, bone marrow, heart, lung, skin, cornea, eye, spinal cord, uterus, intestine, heart valve, bone, cartilage, tendon, ligament, lymphatic vessel, and blood vessel. ### Claim 26 (depends on 24) The mammalian organ or tissue of any one of claim 24, wherein the second mammal is selected from the group consisting of mouse, rat, rabbit, guinea pig, human, cow, pig, horse, goat, and sheep. ### Claim 27 (canceled) ### Claim 28 (depends on 24) The mammalian organ or tissue of claim 24, wherein the first mammal is selected from the group consisting of mouse, rat, rabbit, guinea pig, cow, pig, horse, goat, and sheep. ### Claim 29 (canceled) ### Claim 30 (depends on 24) The mammalian organ or tissue of claim 24, wherein cells of the first mammal comprise a genetic modification at one or more loci, wherein the genetic modification inactivates at least one gene selected from the group consisting of FAH, NKX2.5, TBX5, MEF2C, Osr1, Lhx1, Pax2, Pax8, Wt1, Hox11, Eya1, Six2, Sal11, Etv2, Trox1, Ronx-1, Scl/Tal-1, Lmo-2, Tel, Tek, Sox9, Scleraxis, Pax6, and Rx. ### Claim at position 9 -33. (canceled) ### Claim 34 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises a genetic modification that inactivates Pdx1 and the cells from the second mammal form at least a portion of a pancreas. ### Claim 35 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Runx-1, Scl/Tal-1, Lmo-2, Tel, and/or Tek and the cells from the second mammal form at least a portion of a blood vessel or hematopoietic cell. ### Claim 36 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate FAH and the cells from the second mammal form at least a portion of a liver. ### Claim 37 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Osr1, Lhx1, Pax2, Pax8, Wt1, Hox11, Eya1, Six2, and/or Sal11 and the cells from the second mammal form at least a portion of a kidney. ### Claim 38 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Nkx2.5, Tbx5, and/or Mef2c and the cells from the second mammal form at least a portion of a heart. ### Claim 39 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Pax6 and/or Rx and the cells from the second mammal form at least a portion of an eye. ### Claim 40 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Sox9 and/or Scleraxis and the cells from the second mammal form at least a portion of a cartilage tissue. ### Claim 41 (depends on 24) The mammalian organ of claim 24, wherein the first mammal comprises genetic modifications that inactivate Etv2 and/or Prox1 and the cells from the second mammal form at least a portion of an endothelial or lymphatic vessel. ### Claim at position 18 -69. (canceled) ### Claim 70 A method of making an induced pluripotent stem cell (iPSC) comprising: a) transfecting a population of human foreskin fibroblasts (HFF) with at least three episomal vectors comprising pCXLE-hOCT3/4-shp53-F, pCXLE-hSK, and pCXLE-hUL; b) contacting the transfected population of HFFs with a population of mitotically inactivated mouse embryonic fibroblasts (MEFs); c) contacting the population of HFFs and the population of MEFs with an FAC medium comprising a base media, an FGF, an Activin, and a WNT activator until at least one colony comprising an iPSC has formed; and d) transferring the colony comprising an iPSC to a fresh population of mitotically inactivated MEFs. ### Claim 71 (depends on 70) The method of claim 70, wherein the base media comprises DMEM/F12, Neurobasal medium, N2 supplement, B27 supplement, GlutaMax, non-essential amino acids, beta-mercaptoethanol, and antibiotics. ### Claim 72 (depends on 70) The method of claim 70, wherein the FGF comprises an FGF2. ### Claim 73 (depends on 70) The method of claim 70, wherein the Activin comprises an Activin A. ### Claim 74 (depends on 70) The method of claim 70, wherein the WNT activator comprises a GSK inhibitor. ### Claim 75 (canceled) ### Claim 76 (depends on 70) The method of claim 70, wherein the HFF are transfected with pCXLE-EGFP. ### Claim 77 (depends on 70) The method of claim 70, wherein the FAC medium comprises bovine serum albumin