公开(公告)号：US20160272937A1

公开(公告)日：2016-09-22

申请号：US15030315

申请日：2014-04-14

申请人： THE BRIGHAM AND WOMEN'S HOSPITAL, INC.

发明人： Albert LAM ,  Benjamin FREEDMAN ,  Ryuji MORIZANE ,  Joseph BONVENTRE

IPC分类号： C12N5/0735 ,  A61K35/54

CPC分类号： C12N5/0611 ,  A61K35/54 ,  C12N5/0687 ,  C12N2500/90 ,  C12N2501/115 ,  C12N2501/119 ,  C12N2501/16 ,  C12N2501/385 ,  C12N2501/724 ,  C12N2501/727 ,  C12N2506/02 ,  C12N2506/45

摘要： Described herein are methods related to generating intermediate mesoderm (IM) cells, including using sequential treatment of small molecules and growth factors, and composition produced by the described methods. Using small molecules such as CHIR99021 in combination with FGF2 and RA, efficient differentiation of human pluripotent stem cells (hPSCs) into intermediate mesoderm, such as PAX2+LHX1+ cells, is achieved. The method is extensible different hPSC cell lines and does not require flow sorting. Importantly, resulting PAX2+LHX1+ cells, are capable of WT1 expression and addition of FGF9 and activin, PAX2+LHX1+ cells specifically differentiates cells into SIX2, SALL1, and WT1 expressing cells representative of cap mesenchyme nephron progenitor cells. The described methods and compositions facilitate and improve the directed differentiation of hPSCs into cells of the kidney lineage for the purposes of bioengineering kidney tissue and iPS cell disease modeling.

摘要翻译： 本文描述了与产生中间胚层（IM）细胞相关的方法，包括使用小分子和生长因子的顺序处理以及由所述方法产生的组合物。 使用小分子如CHIR99021与FGF2和RA组合，实现人多能干细胞（hPSC）到中间胚层如PAX2 + LHX1 +细胞的有效分化。 该方法是可扩展的不同的hPSC细胞系，不需要流分选。 重要的是，所得到的PAX2 + LHX1 +细胞能够进行WT1表达，并加入FGF9和激活素，PAX2 + LHX1 +细胞将细胞特异性分化为表达帽间质肾单核祖细胞的SIX2，SALL1和WT1表达细胞。 所描述的方法和组合物促进和改善hPSC到肾脏谱系细胞的定向分化，用于生物工程肾组织和iPS细胞疾病建模。