IntroductIonH arnessing the differentiation potential of pluripotent stem cells could be of great clinical value for treatment of degenerative diseases. in human development, pluripotent stem cells appear transiently in the inner cell mass of the blastocyst, but if isolated and placed under appropriate culture conditions, the embryonic stem (es) cell lines can be established and propagated in vitro seemingly indefinitely. 1 Closely analogous to the es cells are embryonic carcinoma (eC) cells. eC cells are the pluripotent cells of a subset of testicular germ cell nonseminoma tumors, which arise as a consequence of neoplastic transformation of primordial germ cells. 2 numerous cell lines with different degrees of differentiation ability have been established from human teratocarcinomas. one of the best-characterized pluripotent human eC cell lines is ntera2. 3 ntera2 cells share with es cells the expression of pluripotency-associated genes (e.g., OCT4 [officially termed POU5F1] and NANOG), as well as the expression of surface antigens such as glycolipid antigens stage-specific antigen 3 (ssea3) and ssea4 and the proteoglycan antigens tra-1-60 and tra-1-81. 4,5 although ntera2 cells are propagated in vitro in an undifferentiated stem cell state, their differentiation can be induced in response to chemical agents, the best studied of which is all-trans retinoic acid. 6 following the exposure of cells to all-trans retinoic acid, cells lose their characteristic morphology and the expression of key markers of undifferentiated state (oCt4, nanog, ssea3, ssea4, tra-1-60, and tra-1-81) and begin to express a range of new markers, such as ssea1 and a2B5. 4,5,7 the rate of disappearance of markers characteristic of the undifferentiated state differs markedly, with ssea3 disappearing most rapidly. 4,5,7 therefore, ssea3 represents a particularly sensitive marker of undifferentiated stem cell state.the applicability of all-trans retinoic acid in inducing differentiation of eC cells provided a paradigm for the role of small chemicals in controlling the ntera2 stem cell fates. 6,8 subsequently, all-trans retinoic acid also proved useful in inducing differentiation of es cells. 9 the advantage of using small molecules over genetic approaches is that they are easily applied to cells and allow reversible temporal and dosedependent control of protein function. additional compounds that could be used as supplements in the media for directed differentiation and/or tools for dissecting molecular pathways governing stem cell biology would be beneficial. High-content Screening for chemical Modulators of Embryonal carcinoma cell differentiation and SurvivalIvAnA BArBArIc, MArk JonES, dAvId J. HArLEy, PAuL J. GokHALE, and PEtEr W. AndrEWS disentangling the complex interactions that govern stem cell fate choices of self-renewal, differentiation, or death presents a formidable challenge. image-based phenotype-driven screening meets this challenge by providing means for rapid testing of many small molecules simultaneously. pluripotent embry...
The antigenic profile of human embryonic stem (ES) and embryonal carcinoma (EC) cells has served as a key element of their characterization, with a common panel of surface and intracellular markers now widely used. Such markers have been used to identify cells within the 'undifferentiated state', yet it appears that this categorization may be an oversimplification, because a number of sub-states appear to exist within this state. To increase the resolution of the undifferentiated state, we have generated eight novel monoclonal antibodies, all capable of recognizing undifferentiated human ES and EC cells, and herein describe their characterization. The reactivity of these antibodies against a range of cell lines is reported, as well as their developmental regulation, basic biochemistry and reactivity in immunohistochemistry of testicular germ cell tumours. Our data reveal a range of reactivity for all antibodies against both ES and EC cells, suggesting that these markers will afford recognition of unique sub-states within the undifferentiated stem cell compartment.
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