Regulation of human germline commitment by DMRT1 [scRNA-Seq]

Nascent human primordial germ cell-like cells (PGCLCs) can be specified with BMP2 or BMP4 together with SCF and EGF from germline competent embryonic stem cells which are maintained in the presence of inhibitors for GSK3, MEK, p38 and JNK together with FGF2, TGFβ and LIF (4i ESC). Here we discovered that Activin A and retinoic acid treatment after PGC specification induced genes expressed in migrating and gonadal PGCs in vivo, including DMRT1 and CDH5. Ectopic expression of DMRT1 and SOX17 resulted in repression of pluripotent genes as well as induction of DAZL and a subset of mitotic arrest PGC genes. Our findings provide novel insights on the roles of DMRT1 for the transition from nascent PGCs towards germline determination and molecular mechanisms of early human germline development. 10x RNA profiles of 4i ESC, NANOS3+ PGCLCs, DMRT1+ PGCLCs, and d8 DAZL+ PGCLCs.

新生人类原始生殖细胞样细胞（primordial germ cell-like cells, PGCLCs）可由采用GSK3、MEK、p38及JNK抑制剂联合FGF2、TGFβ与LIF进行培养的生殖系潜能胚胎干细胞（4i ESC），经BMP2或BMP4联合SCF与EGF诱导分化获得。本研究发现，在原始生殖细胞特化完成后施加激活素A（Activin A）与视黄酸处理，可诱导体内迁移期及性腺期原始生殖细胞的特征基因表达，其中涵盖DMRT1与CDH5。异位过表达DMRT1与SOX17可抑制多能基因的转录，同时诱导DAZL及部分有丝分裂阻滞期原始生殖细胞特征基因的表达。本研究结果为阐明DMRT1在新生原始生殖细胞向生殖系命运决定的转化过程中的作用，以及早期人类生殖系发育的分子机制提供了全新的研究视角。本数据集包含4i ESC、NANOS3阳性PGCLCs、DMRT1阳性PGCLCs以及d8 DAZL阳性PGCLCs的10x RNA测序谱。