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  • MRPS18-2 protein immortalizes primary rat embryonic fibroblasts and endows them with stem cell-like properties.

MRPS18-2 protein immortalizes primary rat embryonic fibroblasts and endows them with stem cell-like properties.

Proceedings of the National Academy of Sciences of the United States of America (2009-11-12)
Elena Kashuba, Surya Pavan Yenamandra, Suhas Deoram Darekar, Mariya Yurchenko, Vladimir Kashuba, George Klein, Laszlo Szekely
ABSTRACT

We report that the overexpression of mitochondrial ribosomal protein MRPS18-2 (S18-2) can immortalize primary rat embryonic fibroblasts (REFs). The immortalized cells (18IM) lose contact inhibition, form foci, and are capable of anchorage-independent growth. Concurrently, mesodermal markers, such as vimentin, smooth muscle actin, and Fut4, disappear completely. 18IM cells express embryonic stem cell markers, such as SSEA-1, Sox2, and Oct3/4. In confluent cultures, a portion of cells also express ectoderm- and endoderm-specific pan-keratin, ectoderm-specific beta-III-tubulin, mesoderm-specific MHC class II, and become stainable for fat with Oil red O. None of these changes was detected in c-myc+Ha-ras (MR)-transformed cells. In immunodeficient mice, 18IM cells formed small transiently growing tumors that have down-regulated SSEA-1 and showed pan-keratin staining. We conclude that S18-2 can immortalize REFs and induces them to express stem cell traits.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Oil Red O, certified by the Biological Stain Commission
Sigma-Aldrich
GenElute Mammalian Total RNA Miniprep Kit, sufficient for 70 purifications
Sigma-Aldrich
GenElute Mammalian Total RNA Miniprep Kit, sufficient for 350 purifications
Supelco
Oil Red O, BioReagent, suitable for electrophoresis
Sigma-Aldrich
GenElute Mammalian Total RNA Miniprep Kit, sufficient for 10 purifications
Sigma-Aldrich
Enhanced Avian First Strand Synthesis Kit, Components for cDNA synthesis with enhanced AMV reverse transcriptase

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