Genetic markers for engraftment of human cardiac ventricular progenitor cells
First Claim
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1. A method of transplanting engraftable human ventricular progenitor cells (HVPs), the method comprising:
- culturing human cells containing cardiac progenitor cells (CPCs) under conditions causing differentiation into human ventricular progenitor cells (HVPs) by subjecting human pluripotent stem cells to activation of Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5 to thereby obtain a culture of day 5-7 CPCs comprising day 5-7 LIFR+ Islet1+HVPs;
detecting expression on the HVPs of at least one surface marker selected from the group consisting of JAG1, FZD4, FGFR3 and/or TNFSF9 on the day 5-7 HVPs;
detecting expression in the day 5-7 HVPs of at least one engraftment markers;
isolating day 5-7 HVPs that co-express the at least one surface marker and the at least one engraftment markers to thereby isolate engraftable day 5-7 HVPs; and
transplanting the engraftable day 5-7 HVPs into a subject such that the engraftable day 5-7 HVPs form a vascularized, electrically responsive ventricular muscle patch that secretes an extracellular matrix.
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Abstract
The present invention provides genetic markers for identifying engraftable human cardiac ventricular progenitor cells. The engraftment markers of the invention include angiogenic markers and extracellular matrix markers. Human ventricular progenitor cells expressing these markers are capable of forming ventricular tissue in vivo that is vascularized and supported by an extracellular matrix. Methods of engrafting human cardiac ventricular progenitor cells by transplanting into a subject progenitor cells that express the engraftment markers are also provided.
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Citations
38 Claims
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1. A method of transplanting engraftable human ventricular progenitor cells (HVPs), the method comprising:
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culturing human cells containing cardiac progenitor cells (CPCs) under conditions causing differentiation into human ventricular progenitor cells (HVPs) by subjecting human pluripotent stem cells to activation of Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5 to thereby obtain a culture of day 5-7 CPCs comprising day 5-7 LIFR+ Islet1+HVPs;detecting expression on the HVPs of at least one surface marker selected from the group consisting of JAG1, FZD4, FGFR3 and/or TNFSF9 on the day 5-7 HVPs; detecting expression in the day 5-7 HVPs of at least one engraftment markers;
isolating day 5-7 HVPs that co-express the at least one surface marker and the at least one engraftment markers to thereby isolate engraftable day 5-7 HVPs; andtransplanting the engraftable day 5-7 HVPs into a subject such that the engraftable day 5-7 HVPs form a vascularized, electrically responsive ventricular muscle patch that secretes an extracellular matrix. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. A method of identifying engraftable human ventricular progenitor cells (HVPs), the method comprising:
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detecting expression of three or more positive extracellular matrix markers selected from the group consisting of;
FGF10, SMOC1, CCBE1, COL6A6, ADAMTS12, COL19A1, LAMA1, BMP4, FBLN7, FBLN2, NDNF, HTRA1, HAPLN1, EMILIN1, SPOCK3, PODNL1, IHH, ACAN, NID2, COL4A6, LAMC1, FMOD, MUC4, EMID1, HMCN1, NID1, VCAN, CILP2, SOD3, ADAMTS3, ZP3, ANGPTL4, CRTAC1, LTBP4 and FREM1 in the HVPs to thereby identify engraftable HVPs, wherein;the HVPs are obtained from a culture of day 5-7 cardiac progenitor cells (CPCs), wherein the culture of day 5-7 CPCs is obtained by subjecting human pluripotent stem cells to activation of the Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5; and
whereinthe HVPs also express at least one surface marker selected from the group consisting of;
JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9. - View Dependent Claims (26, 27)
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28. A method of identifying engraftable human ventricular progenitor cells (HVPs), the method comprising:
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detecting expression of at least one positive angiogenesis marker selected from the group consisting of PRKD1, CCBE1, PDGFRA, EPHB2, GATA2, NTRK1, PTGIS, BMPER, C1GALT1, MEIS1, TBX1, PKNOX1, ID1, TCF21, HEY1, HOXB3, HGF, IL6, GHRL, IHH, SRPK2, HAND1, AMOT, NRP2, PTEN, SEMA3E, APOLD1, SETD2, DAB2IP, KDR, PGF, EMP2, TAL1, HIPK2, CSPG4, TNFAIP3, NRP1, NFATC4, CDC42, ANGPTL4, BCAS3, HIPK1, NRXN3, FZD5 and HHEX; and
/ordetecting expression of at least one positive extracellular matrix markers selected from the group consisting of SMOC1, CCBE1, COL6A6, ADAMTS12, COL19A1, LAMA1, FBLN7, FBLN2, NDNF, HTRA1, HAPLN1, EMILIN1, SPOCK3, PODNL1, IHH, ACAN, NID2, COL4A6, LAMC1, FMOD, MUC4, EMID1, HMCN1, NID1, VCAN, CILP2, SOD3, ADAMTS3, ZP3, ANGPTL4, CRTAC1, LTBP4 and FREM1 in the HVPs to thereby identify engraftable HVPs, wherein; the HVPs are obtained from a culture of day 5-7 cardiac progenitor cells (CPCs), wherein the culture of day 5-7 CPCs is obtained by subjecting human pluripotent stem cells to activation of the Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5; and
whereinthe HVPs also express at least one surface marker selected from the group consisting of;
JAG1, FZD4, LIFR and/or TNFSF9.- View Dependent Claims (29, 30, 31, 32, 33)
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34. A method of identifying engraftable human ventricular progenitor cells (HVPs), the method comprising:
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detecting expression of ten or more positive angiogenic markers in the HVPs to thereby identify engraftable HVPs, wherein; the HVPs are obtained from a culture of day 5-7 cardiac progenitor cells (CPCs), wherein the culture of day 5-7 CPCs is obtained by subjecting human pluripotent stem cells to activation of the Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5; and
whereinthe HVPs also express at least one surface marker selected from the group consisting of;
JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9. - View Dependent Claims (35)
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36. A method of identifying engraftable human ventricular progenitor cells (HVPs), the method comprising:
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detecting expression of five or more positive extracellular matrix markers in the HVPs to thereby identify engraftable HVPs, wherein; the HVPs are obtained from a culture of day 5-7 cardiac progenitor cells (CPCs), wherein the culture of day 5-7 CPCs is obtained by subjecting human pluripotent stem cells to activation of the Wnt/β
-catenin signaling on day 0, followed by inhibition of Wnt/β
-catenin signaling from day 3 to day 5; and
whereinthe HVPs also express at least one surface marker selected from the group consisting of;
JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9. - View Dependent Claims (37, 38)
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Specification