Journal: Stem Cells Translational Medicine

Article Title: Putative Immunogenicity Expression Profiling Using Human Pluripotent Stem Cells and Derivatives

doi: 10.5966/sctm.2014-0117

Figure Lengend Snippet: ZG16 expression increased significantly on differentiation of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) into clinically relevant cell derivatives. (A): HORMAD1 expression in hESC and hiPSC derivatives was very low and heterogeneous (top). ZG16 expression analysis resulted in a significant increase across all four lines on differentiation (bottom). ∗, p < .05 for HORMAD1 expression for fibroblast-2 line compared with all other PSC derivatives. 𝔛∗∗, p < .05 for human fetal OPCs compared with other OPCs derived from all lines. ∗∗∗, p < .05 for each PSC line undifferentiated compared with differentiated. Error bars represent ±SD. The y-axis is a logarithmic scale, with display units of 100. All polymerase chain reactions were run in triplicate. (B): Gene expression analysis for ZG16 was assessed in hiPSC1- and hiPSC3-derived hepatocytes. The significant increase in ZG16 expression seen during hepatocyte differentiation from control iPSCs is reproducible across both iPS1 and iPS3 lines. A, B, and C listed in the cell line annotations indicate the duplicate differentiation protocol across three replicates. ∗, p < .05 for iPS1-A, -B, and -C differentiated replicates compared with iPS3-A, -B, and -C differentiated replicates; ∗∗, p < .05 for iPS3-D compared with iPS3-A, -B, and -C differentiated replicates; and ∗∗∗, p < .05 for iPS3-A–D compared with iPS3-B–D. ZG16 y-values are in units of hundreds. Error bars represent ±SD. Abbreviations: Card., cardiomyocytes; Card.-A, adult cardiomyocytes; dif, differentiation; ES, human embryonic stem cell line; Hep., hepatocytes; Hep.-A, adult hepatocytes; OPCs, oligodendrocyte progenitor cell; PSC, pluripotent stem cell.

Article Snippet: LAVIV adult human skin-derived dermal fibroblasts were obtained from a 4-mm skin punch biopsy, as described in the Isolagen Standardized Manufacturing Process EX-GTR-110, version 00 (Fibrocell Science).

Techniques: Expressing, Derivative Assay, Gene Expression, Control

Journal: Cell Division

Article Title: Epitope of titin A-band-specific monoclonal antibody Tit1 5 H1.1 is highly conserved in several Fn3 domains of the titin molecule. Centriole staining in human, mouse and zebrafish cells

doi: 10.1186/1747-1028-7-21

Figure Lengend Snippet: Immunofluorescence co-localization of the MAb Tit1 5 H1.1 target antigen (titin) with respect to actin filaments and some centrosome/centriole proteins. (A) Fibroblasts of the human adult fibroblast cell-line SA-54 were cultured for 3 weeks, fixed with 4% PFA,permeabilized, and double labeled with Alexa Fluor 488 conjugated to Phalloidin for F-actin (green) and with the MAb Tit1 5 H1.1 primary antibody for titin (specific staining detected with Alexa Fluor 594 conjugated to goat anti-mouse IgG secondary antibody (red). (B,C) Mouse embryonal fibroblasts (MEF7) cultured for 3 days, fixed with 4% PFA, permeabilized, and double labeled with Alexa Fluor 488 conjugated to Phalloidin for F-actin (green) and with the MAb Tit1 5 H1.1 primary antibody for titin (specific staining with Alexa Fluor 594, red). (D) Bowes melanoma cells cultured for 2 days, fixed with 4% PFA, permeabilized, and double labeled with MAb Tit1 5 H1.1 for titin (red) and with goat anti-human ninein for ninein (green), and (E) with MAb Tit1 5 H1.1 for titin (red) and with rabbit anti-human γ-tubulin for γ-tubulin (green). (F) Fibroblasts of the human adult fibroblast cell-line SA-54 were cultured for 3 weeks, fixed with 4% PFA, permeabilized, and double labeled with MAb Tit1 5 H1.1 for titin (red) and with rabbit anti-human pericentrin for pericentrin (green). Note the individual staining of the target antigens of all antibodies with centrioles/centrosomes. Arrows show centrosome/centriole staining. Cell nuclei were stained blue with DAPI (obj. 100x).

Article Snippet: In this study an original normal human adult skin cell culture SA-54 (developed by LabAs Ltd.) and the commercial cell-line human melanoma Bowes were used.

Techniques: Immunofluorescence, Cell Culture, Labeling, Staining