Control iPSC lines

Characterization of iMA-1L iPSCs cell line

Control iPSC line – iMA-1L
The induced pluripotent stem cell (iPSC) line iMA-1L was generated from human embryonic dermal fibroblasts using episomal vectors expressing pluripotency factors. The episomes were delivered into the cells by nucleofection (NHDF Nucleofector Kit, Lonza). iMA-1L intensively grew in the pluripotent stem cell-like colonies, had dense intercellular contacts, large nuclear-cytoplasmic ratio and expressed endogenous alkaline phosphatase. Immunofluorescent analysis for pluripotency markers showed the expression of OCT4, SOX2, NANOG, and TRA-1-60. iMA-1L has a normal karyotype 46: XY at both early (6) and late (22-24) passages. Immunofluorescent staining showed that iMA-1L can be differentiated into the cells of three germlines. The characteristics of this line was published in Supplementary Information in (Grigor’eva et al., 2020 https://doi.org/10.1007/s10616-020-00406-7).

Characterization of iMA-1L iPSCs cell line
A. Morphology of iPSC colony. B. Histochemical detection of endogenous alkaline phosphatase activity in iMA-1L cells. C. Karyotype (G-banding) (46, XY) of iMA-1L. D. Immunofluorescent analysis for the pluripotency markers OCT4 (red signal), SOX2 (green signal), NANOG (green signal) and TRA-1-60 (red signal) in iPSC line. E. Immunofluorescent analysis for markers of endoderm: GATA6 (green signal); mesoderm: αSMA (red signal), COLLAGEN I (green signal), COLLAGEN IV (red signal), FIBRONECTIN (red signal), VIMENTIN (red signal); ectoderm: GFAP (green signal), NF200 (green signal), SOX1 (red signal). Nuclei were stained with DAPI (blue signal). The scale bar A, D, E (endoderm and mesoderm) – 100 μm, B and E (ectoderm) – 500 μm.

Control iPSC line – iMA-1L
The induced pluripotent stem cell (iPSC) line iMA-1L was generated from human embryonic dermal fibroblasts using episomal vectors expressing pluripotency factors. The episomes were delivered into the cells by nucleofection (NHDF Nucleofector Kit, Lonza). iMA-1L intensively grew in the pluripotent stem cell-like colonies, had dense intercellular contacts, large nuclear-cytoplasmic ratio and expressed endogenous alkaline phosphatase. Immunofluorescent analysis for pluripotency markers showed the expression of OCT4, SOX2, NANOG, and TRA-1-60. iMA-1L has a normal karyotype 46: XY at both early (6) and late (22-24) passages. Immunofluorescent staining showed that iMA-1L can be differentiated into the cells of three germlines. The characteristics of this line was published in Supplementary Information in (Grigor’eva et al., 2020 https://doi.org/10.1007/s10616-020-00406-7).

Characterization of the ICGi021-A and ICGi022-A iPSC lines

Control iPSC lines – ICGi021-A and ICGi022-A
ICGi021-A and ICGi022-A iPSC lines were obtained by reprogramming peripheral blood mononuclear cells from two healthy women of the Siberian population using episomal non-integrating vectors expressing Yamanaka factors. iPSC lines expressed pluripotency markers, have a normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers. Clinical exome sequencing data of the original biosamples of the donors are available in the NCBI SRA database (SRR11413028 and SRR11413027). The generated cell lines are useful as “healthy” control in hereditary disease studies, and for basic research of cellular and molecular mechanisms of pluripotency and differentiation.

Characterization of the ICGi021-A and ICGi022-A iPSC lines
A. Morphology of iPSC colony. B. Alkaline phosphatase staining. C. Immunofluorescence staining for pluripotency markers SSEA-4, OCT4, SOX2 and TRA-1-60. D. Quantitative analysis of pluripotency marker expression (NANOG, OCT4, SOX2) by qPCR. E. Elimination of the episomal reprogramming vectors confirmed by PCR. F. Mycoplasma testing by PCR – Negative. G. In vitro differentiation. Immunofluorescence staining for differentiation markers: NF200 and TUBB3 (ectoderm); aSMA and COLL I (mesoderm); HNF3b and TBX3 (endoderm).

Control iPSC lines – ICGi021-A and ICGi022-A
ICGi021-A and ICGi022-A iPSC lines were obtained by reprogramming peripheral blood mononuclear cells from two healthy women of the Siberian population using episomal non-integrating vectors expressing Yamanaka factors. iPSC lines expressed pluripotency markers, have a normal karyotype and demonstrated the ability to differentiate into derivatives of the three germ layers. Clinical exome sequencing data of the original biosamples of the donors are available in the NCBI SRA database (SRR11413028 and SRR11413027). The generated cell lines are useful as “healthy” control in hereditary disease studies, and for basic research of cellular and molecular mechanisms of pluripotency and differentiation.