Figure 1

Infected hESCs cured with PlasmocinTM maintain their morphology, stem cell markers expression and karyotype.Mycoplasma sp. infected HUES-5 884 (H5 884) cells were treated with Plasmocin 25 µg/ml during 14 days (Curative treatment) and: (A) Mycoplasma sp. infection was followed by genomic DNA PCR analysis at day 0, 7 and 14 after antibiotic addition (lanes 4, 2 and 3), in non infected parental H5 line (lane 1) and in cured cells after 6 month post-treatment (lane 7) Control +: Mycoplasmaorale genomic DNA (lanes 5 and 9). Control -: H2O (lanes 6 and 8); (B) Cured H5 884 cells were photographed using an inverted microscope in order to compare to HUES-5 (H5) colony morphology. Figure shows representative bright field photomicrographs. Scale bars = 100 µm; (C) H5 and cured H5 884 cells were grown on MatrigelTM coated plates until confluence and stained with primary antibodies that recognize stem cell markers. Figure shows representative fluorescent photomicrographs of hESCs immunostained with SSEA-4, TRA-1-60, TRA-1-81, Nanog and Oct-4. Scale bars = 100 µm; (D) H5 884 mRNA levels of oct-4 and nanog were analyzed by RT-Real Time PCR after 14 days of curative treatment and compared to H5 levels. Rpl7 expression was used as normalizer. Graph shows mRNA fold induction relative to human fibroblasts (HF). The mean ± S.E. from three independent experiments are shown. * = p<0.05; (E) Karyotype analysis of cured H5 884 cells. A representative GTG-banded metaphase spread is shown. Abbreviations: SSEA-4, Stage-Specific Embryonic Antigen; TRA-1-60, Tumor Rejection Antigen 1-60; TRA-1-81, Tumor Rejection Antigen 1-81; Oct-4, Octamer 4.

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