Improved generation of C57BL/6J mouse embryonic stem cells in a defined serum-free media

C57BL/6 is a well-characterized mouse strain that is used extensively for immunological and neurological research. The establishment of C57BL/6 ES cell lines has facilitated the study of gene-altered mice in a pure genetic background - however, relatively few such lines exist. Using a defined media supplement, knockout serum replacement (KSR) with knockout DMEM (KSR-KD- MEM), we find that we can readily establish ES cell lines from blastocysts of C57BL/6J mice. Six lines were established, all of which were karyotypically normal and could be maintained in the undifferentiated state on mouse embryonic fibroblast (MEF) feeders. One line was further tested and found to be karyotypically stable and germline competent, both prior to manipulation and after gene targeting. For this cell line, efficiencies of cell cloning and chimera generation were greater when maintained in KSR-KDMEM. Our work suggests that the use of defined serum-free media may facilitate the generation of ES cells from inbred mouse strains. genesis Published 2004 Wiley-Liss, Inc.(dagger)

C57BL/6是一种特征明确的小鼠品系，广泛用于免疫学和神经学研究。C57BL/6胚胎干细胞（ES细胞）系的建立促进了在纯遗传背景下对基因改变小鼠的研究——然而，此类细胞系相对较少。我们使用一种特定的培养基补充剂，即含有无血清替代物（KSR）的无血清DMEM（KSR - KD - MEM），发现能够很容易地从C57BL/6J小鼠的囊胚中建立ES细胞系。建立了6个细胞系，它们的染色体核型均正常，并且可以在小鼠胚胎成纤维细胞（MEF）饲养层上维持未分化状态。对其中一个细胞系进行了进一步测试，发现在基因操作之前和基因打靶之后，其染色体核型稳定且具有生殖系能力。对于这个细胞系，在KSR - KDMEM中维持时，细胞克隆和嵌合体产生的效率更高。我们的工作表明，使用特定的无血清培养基可能有助于从近交系小鼠品系中产生ES细胞。《起源》，2004年威利 - 利斯公司出版（†）