Ritner Carissa, Bernstein Harold S
Cardiovascular Research Institute, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, CA, USA.
J Vis Exp. 2010 Aug 1(42):2036. doi: 10.3791/2036.
Human embryonic stem cells (hESCs) have an unlimited capacity for self-renewal, and the ability to differentiate into cells derived from all three embryonic germ layers. Directed differentiation of hESCs into specific cell types has generated much interest in the field of regenerative medicine (e.g., (2-5)), and methods for determining the in vivo fate of selected or manipulated hESCs are essential to this endeavor. We have adapted a highly efficient teratoma formation assay for this purpose. A small number of specifically selected hESCs is mixed with undifferentiated wild type hESCs and Phaseolus vulgaris lectin to form a cell pellet. This is grafted beneath the kidney capsule in an immunodeficient mouse. As few as 2.5 x 10(5) hESCs are needed to form a 16 cm(3) teratoma within 8-12 weeks. The fate of the originally selected hESCs can then be determined by immunohistochemistry. This method provides a valuable tool for characterizing tissue-specific reagents for cell-based therapy.
人类胚胎干细胞（hESCs）具有无限的自我更新能力，以及分化为源自所有三个胚胎胚层的细胞的能力。将hESCs定向分化为特定细胞类型已在再生医学领域引起了广泛关注（例如，(2 - 5)），而确定所选或经操作的hESCs在体内命运的方法对于这项工作至关重要。为此，我们采用了一种高效的畸胎瘤形成试验。将少量经过特定选择的hESCs与未分化的野生型hESCs和菜豆凝集素混合形成细胞团块。将其移植到免疫缺陷小鼠的肾包膜下。在8 - 12周内形成一个16 cm³ 的畸胎瘤仅需要低至2.5×10⁵个hESCs。然后可以通过免疫组织化学确定最初所选hESCs的命运。该方法为表征用于细胞治疗的组织特异性试剂提供了一种有价值的工具。
