title: Mycoplasma contamination of murine embryonic stem cells affects cell parameters, germline transmission and chimeric progeny.
abstract: Murine embryonic stem cells (mESCs) inoculated at passage P13 with the mycoplasma species M. hominis, M. fermentans and M. orale and cultured over 20 passages showed reduced growth rate and viability (P < 0.0001) compared to control mESCs. Spectral karyotypic analysis of mycoplasma-infected mESCs showed a number of non-clonal chromosomal aberrations which increased with the duration of infection. The differentiation status of the infected mESCs was most affected at passage P13+6 where the infection was strongest and 46.3% of the mESCs expressed both POU5F1 and SSEA-1 markers whereas 84.8% of control mESCs expressed both markers. The percentage of germline chimeras from mycoplasma-infected mESCs was examined after blastocyst injection and embryo transfer to suitable recipients at different passages and, compared to the respective control group, was most affected at passage P13+5 (50% vs. 90%; P < 0.07). Further reductions were obtained at the same passage in the percentage of litters born (50% vs. 100%; P < 0.07) and in the percentage of pups born (22% vs. 45%; P < 0.001). Thirty three chimeras (39.8%) obtained from blastocyst injection with mycoplasma-infected mESCs showed reduced body weight (P < 0.0001), nasal discharge, osteoarthropathia, and cachexia. Flow cytometric analysis of plasma from chimeras produced with mycoplasma-infected mESCs revealed statistically significant differences in the proportions of T-cells and increased levels of IgG1 (P < 0.001), IgG2a (P < 0.05) and IgM (P < 0.05), anti-DNA antibodies (P < 0.05) and rheumatoid factor (P < 0.01). The present data indicate that mycoplasma contamination of mESCs affects various cell parameters, germline transmission, and postnatal development of the resulting chimeras.
authors.affiliation: Department of Comparative Medicine, Helmholtz Center Munich, German Research Center for Environmental Health, Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany.
authors.forename: Kyriaki
authors.initials: K
authors.lastname: Markoullis
authors.forename: Diana
authors.initials: D
authors.lastname: Bulian
authors.forename: Gabriele
authors.initials: G
authors.lastname: Hölzlwimmer
authors.forename: Leticia
authors.initials: L
authors.lastname: Quintanilla-Martinez
authors.forename: Katrin-Janine
authors.initials: KJ
authors.lastname: Heiliger
authors.forename: Horst
authors.initials: H
authors.lastname: Zitzelsberger
authors.forename: Hagen
authors.initials: H
authors.lastname: Scherb
authors.forename: Josef
authors.initials: J
authors.lastname: Mysliwietz
authors.forename: Cord C
authors.initials: CC
authors.lastname: Uphoff
authors.forename: Hans G
authors.initials: HG
authors.lastname: Drexler
authors.forename: Thure
authors.initials: T
authors.lastname: Adler
authors.forename: Dirk H
authors.initials: DH
authors.lastname: Busch
authors.forename: Jörg
authors.initials: J
authors.lastname: Schmidt
authors.forename: Esther
authors.initials: E
authors.lastname: Mahabir
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001755:Blastocyst / Q000382:microbiology / Q000502:physiology; D002454:Cell Differentiation; D002470:Cell Survival; D002678:Chimera / Q000382:microbiology / Q000502:physiology*; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000382:microbiology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005854:Germ Cells / Q000502:physiology*; D007074:Immunoglobulin G / Q000378:metabolism; D007621:Karyotyping; D008297:Male; D051379:Mice; D009174:Mycoplasma / Q000302:isolation & purification / Q000502:physiology*; D011247:Pregnancy
chemical_list: D015415:Biomarkers; D007074:Immunoglobulin G