Isolation of human embryonic stem cell-derived teratomas for the assessment of pluripotency.
Isolation of human embryonic stem cell-derived teratomas for the assessment of pluripotency.
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DOI：
10.1002/9780470151808.sc01b04s3
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发表时间：
2007-10-01
影响因子：
--
通讯作者：
Ahrlund-Richter, Lars
中科院分区：
文献类型：
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作者：
Gertow, Karin;Przyborski, Stefan;Ahrlund-Richter, Lars
本单元描述了如何评估人胚胎干细胞（hESC）的发育潜能，通过异种移植到免疫缺陷小鼠诱导畸胎瘤形成的协议。hESC可以注射到睾丸被膜下，或者可选地注射到肾脏中或皮下。从移植的hESC发育的畸胎瘤被手术切除，在甲醛中固定，并石蜡包埋。对畸胎瘤中的组织进行组织学分析，以确定hESC是否具有多能性并形成源自所有三个胚胎种系（外胚层、中胚层和内胚层）的组织。畸胎瘤也可以固定在布因氏或冷冻切片进行分析，并可以通过免疫组织化学分析组织标记物。本单元包括这些程序的方法。
This unit describes protocols on how to assess the developmental potency of human embryonic stem cells (hESCs) by performing xenografting into immunodeficient mice to induce teratoma formation. hESCs can be injected under the testis capsule, or alternatively into the kidney or subcutaneously. Teratomas that develop from grafted hESCs are surgically removed, fixed in formaldehyde, and paraffin embedded. The tissues in the teratoma are analyzed histologically to determine whether the hESCs are pluripotent and form tissues derived from of all three embryonic germ layers (ectoderm, mesoderm, and endoderm). Teratomas can also be fixed in Bouin's or cryosectioned for analysis, and they can be analyzed by immunohistochemistry for tissue markers. Methods for these procedures are included in this unit.