INVESTIGADORES
ROMORINI Leonardo
artículos
Título:
Effect of antibiotics against Mycoplasma sp. on human embryonic stem cells
Autor/es:
LEONARDO ROMORINI; DIEGO A. RIVA; CAROLINA BLUGUERMANN; GUILLERMO A. VIDELA RICHARDSON; MARÍA E. SCASSA; GUSTAVO E. SEVLEVER; SANTIAGO G. MIRIUKA
Revista:
PLOS ONE
Editorial:
PUBLIC LIBRARY SCIENCE
Referencias:
Lugar: San Francisco; Año: 2013 p. 1 - 15
ISSN:
1932-6203
Resumen:
Human embryonic stem cells (hESCs) are self-renewing pluripotent cells that can
differentiate into specialized cells and hold great promise as models for human
development and disease studies, cell-replacement therapies, drug discovery and in
vitro cytotoxicity tests. The culture and differentiation of these cells are both complex
and expensive, so it is essential to extreme aseptic conditions. hESCs are susceptible
to Mycoplasma sp. infection, which is hard to detect and alters stem cell-associated
properties. The purpose of this work was to evaluate the efficacy and cytotoxic effect of
PlasmocinTM and Ciprofloxacin (specific antibiotics used for Mycoplasma sp.
eradication) on hESCs. Mycoplasma sp. infected HUES-5 884 (H5 884, stable hESCs
H5-Brachyury promoter-GFP line) cells were effectively cured with a 14 days
PlasmocinTM 25 ug/ml treatment (curative treatment) while maintaining stemness
characteristic features. Furthermore, cured H5 884 cells exhibit the same karyotype as
the parental H5 line and expressed GFP, through up-regulation of Brachyury promoter,
at day 4 of differentiation onset. Moreover, H5 cells treated with Ciprofloxacin 10 ug/ml
for 14 days (mimic of curative treatment) and H5 and WA09 (H9) hESCs treated with
PlasmocinTM 5 ug/ml (prophylactic treatment) for 5 passages retained hESCs
features, as judged by the expression of stemness-related genes (TRA1-60, TRA1-81,
SSEA-4, Oct-4, Nanog) at mRNA and protein levels. In addition, the presence of
specific markers of the three germ layers (Brachyury, Nkx2.5 and cTnT: mesoderm;
AFP: endoderm; Nestin and Pax-6: ectoderm) was verified in in vitro differentiated
antibiotic-treated hESCs. In conclusion, we found that PlasmocinTM and Ciprofloxacin
do not affect hESCs stemness and pluripotency nor cell viability. However, curative
treatments slightly diminished cell growth rate. This cytotoxic effect was reversible as
cells regained normal growth rate upon antibiotic withdrawal.