公开(公告)号：US20160215263A1

公开(公告)日：2016-07-28

申请号：US14915992

申请日：2014-09-12

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon KELLER ,  Alec Drake WITTY ,  Steven James KATTMAN

IPC: C12N5/077 ,  G01N33/50

Abstract: Provided are methods and products for obtaining cardiovascular lineage cells from hPSCs. The method for obtaining a cardiomyocyte lineage or an epicardial lineage cell population from human pluripotent stem cells (hPSCs) comprises one or more of the following steps: (a) contacting BMP component primed hPSCs with a cardiovascular mesoderm programming cocktail suitable for inducing the hPSCs to differentiate to a cardiovascular mesoderm cell population under conditions suitable for the programming cocktail to penetrate the hPSCs and culturing the contacted hPSCs for a period of time to generate a KDR+ and PDGFRalpha+ cardiovascular mesoderm cell population; (b) contacting the cardiovascular mesoderm cell population with a cardiovascular progenitor specification cocktail suitable to specify a NKX2-5+ or WT1+ cardiovascular progenitor cell population under conditions suitable for the specification cocktail to penetrate the cardiovascular mesoderm cell population and culturing the contacted cardiovascular mesoderm cell population for a period of time to generate a NKX2-5+ or WT1+ cardiovascular progenitor cell population; and (d) contacting the cardiovascular progenitor cell population with a maturation cocktail under conditions suitable for the maturation cocktail to penetrate the cardiovascular progenitor cell population and culturing the contacted cardiovascular progenitor population for a period of time to produce a cardiovascular population optionally cardiomyocyte lineage cells expressing cardiac troponin T (cTnT) and/or SIRPA and/or epicardial lineage cells expressing WT1.

Abstract translation: 提供从hPSC获得心血管谱系细胞的方法和产品。 用于从人多能干细胞（hPSC）获得心肌细胞谱系或心外膜谱系细胞群的方法包括一个或多个以下步骤：（a）使BMP组分引发的hPSC与适合诱导hPSC的心血管中胚层编程混合物接触 在适合编程混合物的条件下分化为心血管中胚层细胞群体以渗透hPSC并培养接触的hPSC一段时间以产生KDR +和PDGFRα+心血管中胚层细胞群; （b）使心血管中胚层细胞群体与适合于规范混合液以渗透心血管中胚层细胞群体并培养接触的心血管中胚层细胞的条件下，适用于指定NKX2-5 +或WT1 +心血管祖细胞群的心血管祖细胞规格混合物接触 人口一段时间以产生NKX2-5 +或WT1 +心血管祖细胞群体; 和（d）在适合于成熟混合物的条件下将心血管祖细胞群与成熟混合物接触以渗透心血管祖细胞群体并培养接触的心血管祖细胞群体一段时间以产生心血管群体，任选地表达心肌细胞谱系细胞 心肌肌钙蛋白T（cTnT）和/或表达WT1的SIRPA和/或心外膜谱系细胞。