Qualification of Embryonal Carcinoma 2102Ep As a Reference for Human Embryonic Stem Cell Research

Abstract

As the number of human embryonic stem cell (hESC) lines increases, so does the need for systematic evaluation of each line's characteristics and potential. Comparisons between lines are complicated by variations in culture conditions, feeders, spontaneous differentiation, and the absence of standardized assays. These difficulties, combined with the inability of most labs to maintain more than a few lines simultaneously, compel the development of reference standards to which hESC lines can be compared. The use of a stable cell line as a reference standard offers many advantages. A line with a relatively unchanging hESC-like gene and protein expression pattern could be a positive control for developing assays. It can be used as a reference for genomics or proteomics studies, especially for normalizing results obtained in separate laboratories. Such a cell line should be widely available without intellectual property restraints, easily cultured without feeders, and resistant to spontaneous changes in phenotype. We propose that the embryonal carcinoma (EC) line 2102Ep meets these requirements. We compared the protein, gene, and microRNA expression of this cell line with those of hESC lines and alternative reference lines such as the EC line NTERA-2 and the karyotypically abnormal hESC line BG01V. The overall expression profiles of all these lines were similar, with exceptions reflecting the germ cell origins of EC. On the basis of global gene and microRNA expression, 2102Ep is somewhat less similar to hESC than the alternatives; however, 2102Ep expresses more hESC-associated microRNAs than NTERA-2 does, and fewer markers of differentiated fates. STEM CELLS 2007;25:437-446

Concise Review: MicroRNA Expression in Multipotent Mesenchymal Stromal Cells

“…Northern blot and cloning analysis identified several microRNAs in human ESC, several of which were identical to microRNAs previously reported in mouse ESC [143]. We recently reported that several microRNAs are highly expressed across multiple hESC lines [144]. Furthermore, as these cells differentiate, the microRNA profiles change significantly.…”

“…Using Northern blot analysis, cloning, or array methods, a set of microRNAs specific to pluripotent cell types, such as mouse ES cells [35], mouse or human embryonic carcinoma cells [22], or human ESC [32,[143][144][145], has been identified. Although these studies could not yet ascribe regulatory functions to these microRNA populations, the unique presence of these populations in stem cells and their disappearance during differentiation suggest roles in maintaining "stemness" by suppressing pluripotency and/or restricting cell differentiation.…”

Cytoplasmic p21 expression levels determine cisplatin resistance in human testicular cancer

“…(B) Enhanced p21 levels in Tera 24 hours after treatment with synthetic anti-miR-106b seed family members. Synthetic anti-miR-220 was used as negative control, since miR-220 expression was not detectable in TC (32). (C) Differences in p21 enhancement in EC cells 24 hours after synthetic anti miR-17-5p treatment.…”

Characterization of microRNAs Involved in Embryonic Stem Cell States

“…Previously, miRNA expression has been examined in murine and human ES cells using cloning, microarray, quantitative polymerase chain reaction (PCR), and deep sequencing technologies on selected cell lines [20][21][22][23][24][25][26][27][28][29]. These studies typically involve embryoid body (EB) formation as an intermediate step, and the differentiation protocols extend for several weeks.…”