Methods Mol Biol. 2011;767:231-41. doi: 10.1007/978-1-61779-201-4_17.
The teratoma assay: an in vivo assessment of pluripotency.
Source
Center for Department of Applied Technology Development, Beckman Research Institute, City of Hope, Duarte, CA, USA. robin@primogenix.com
Abstract
A teratoma is a nonmalignant tumor comprised of a disorganized mixture of cells and small foci of tissue comprised of cells from all three of the embryonic germ-layers. By definition, a cell is pluripotent if it can differentiate into cells derived from all three of the embryonic germ-layers: ectoderm, mesoderm, and endoderm. In the teratoma assay, putative pluripotent stem cells (PSCs) are implanted into an immune-compromised mouse where they may proliferate and differentiate to form a teratoma. The PSCs grow at the implantation site supported by a complex mixture of factors from the local milieu, as well as circulating factors that are vital components of normal mammalian physiology. After a predetermined time of 6-12 weeks or when the tumor has reached sufficient size, it is removed and subjected to histopathological analysis. The teratoma may be further processed by immunocytochemistry and gene expression profiling. This chapter describes methods to generate teratomas through the implantation of putative PSC lines in the SCID mouse. Implantation at the following sites is described: (1) intramuscular, (2) subcutaneous, (3) under the testis capsule, and (4) under the kidney capsule.
Methods Mol Biol. 2011;690:67-80. doi: 10.1007/978-1-60761-962-8_4.
Functional assays for human embryonic stem cell pluripotency.
Source
Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada.
Abstract
Realizing the potential that human embryonic stem cells (hESCs) hold, both for the advancement of biomedical science and the development of new treatments for many human disorders, will be greatly facilitated by the introduction of standardized methods for assessing and altering the biological properties of these cells. The 7-day in vitro alkaline phosphatase colony-forming cell (AP(+)-CFC) assay currently offers the most sensitive and specific method to quantify the frequency of undifferentiated cells present in a culture. In this regard, it is superior to any phenotypic assessment protocol. The AP(+)-CFC assay, thus, provides a valuable tool for monitoring the quality of hESC cultures, and also for evaluating quantitative changes in pluripotent cell numbers following manipulations that may affect the self-renewal and differentiation properties of the treated cells. Two other methods routinely used to evaluate hESC pluripotency involve either culturing the cells under conditions that promote the formation of nonadherent differentiating cell aggregates (termed embryoid bodies), or transplanting the cells into immunodeficient mice to obtain teratomas containing differentiated cells representative of endoderm, mesoderm, and ectoderm lineages.
PLoS One. 2012;7(9):e45532. doi: 10.1371/journal.pone.0045532. Epub 2012 Sep 25.
Standardization of the teratoma assay for analysis of pluripotency of human ES cells and biosafety of their differentiated progeny.
Gropp M, Shilo V, Vainer G, Gov M, Gil Y, Khaner H, Matzrafi L, Idelson M, Kopolovic J, Zak NB, Reubinoff BE.
Source
The Hadassah Human Embryonic Stem Cell Research Center, The Goldyne Savad Institute of Gene Therapy and Department of Gynecology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
The terminology of teratocarcinomas and teratomas.
Lensch MW, Ince TA.
Nat Biotechnol. 2007 Nov;25(11):1211; author reply 1211-2.
Tang C, Lee AS, Volkmer JP, Sahoo D, Nag D, Mosley AR, Inlay MA, Ardehali R, Chavez SL, Pera RR, Behr B, Wu JC, Weissman IL, Drukker M.
Nat Biotechnol. 2011 Aug 14;29(9):829-34. doi: 10.1038/nbt.1947.
Nat Biotechnol. 2011 Sep 8;29(9):803-5. doi: 10.1038/nbt.1974.
Toward safer regenerative medicine.
Comment on
- The following popper user interface control may not be accessible. Tab to the next button to revert the control to an accessible version.Destroy user interface controlAn antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells. [Nat Biotechnol. 2011]
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