Definitive Endoderm Differentiation of BGV Human Embryonic Stem Cells【DOC精选】.docxVIP

下载本文档

3
0
约 6页
2017-10-13 发布于江苏
举报
版权申诉

Definitive Endoderm Differentiation of BGV Human Embryonic Stem Cells【DOC精选】.docx

1、本文档共6页，可阅读全部内容。
2、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。
3、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
5、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
6、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
7、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
8、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

Definitive Endoderm Differentiation of BG01V Human Embryonic Stem Cells This protocol is designed for use with BG01V human embryonic stem cells grown in Mouse Embryonic Fibroblast (MEF) Conditioned Media (AR005). If using different cell lines or growth media, the protocol below may need to be modified. The quality of the human pluripotent cells used in the differentiation is critical. Use of suboptimal quality or very high passage pluripotent cells can result in decreased differentiation efficiency and/or increased cell death. Supplies Required Reagents Supplied in the Human Pluripotent Cell-derived Endoderm Differentiation Kit (Catalog # SC019). ? Endoderm Base Media Supplement ? FGF basic ? Activin A ? Wnt-3a Additional Reagents Required ? RPMI Medium 1640 (Invitrogen, Catalog # 11875 or equivalent) ? DMEM/F-12 (Invitrogen, Catalog # 10565 or equivalent) ? GlutaMAX? (Invitrogen, Catalog # 35050-079 or equivalent) ? Penicillin-Streptomycin (Sigma, Catalog # P4333 or equivalent) ? Phosphate-Buffered Saline (PBS) (Irvine Scientific, Catalog # 9242 or equivalent) ? Cultrex? PathClear?Basement Membrane Extract Reduced Growth Factor (RD Systems, Catalog # 3433-005-02 or equivalent) ? MEF Conditioned Media (RD Systems, Catalog # AR005 or equivalent) ? Trypan Blue Stain (Invitrogen, Catalog # 15250-061 or equivalent) ? Accutase? (Innovative Cell Technologies, Catalog # AT104 or equivalent) ? BSA, very low endotoxin (Millipore, Catalog # 81-068-3 or equivalent) ? Sterile, deionized water Materials ? Human pluripotent stem cells (ATCC, Catalog # SCRC-2002 or equivalent) ? 60 mm tissue culture dishes (Falcon, Catalog # 353002 or equivalent) ? 24-well culture plates (Corning Costar, Catalog # 3526 or equivalent) ? 15 mL centrifuge tubes (Corning Costar, Catalog # 430052 or equivalent) ? 50 mL centrifuge tubes (Corning Costar, Catalog # 430290 or equivalent) ? 0.2 μm syringe filter (PALL Corporation, Catalog # 4187 or equivalent) ? 0.2 μm, 500 mL filter units (Nalgene, Catalo