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Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Oct;29(10):1094-7.

[The stability of synthesized EGFP mRNA in vitro transfected into human umbilical cord mesenchymal stem cells].

[Article in Chinese]

Author information

  • 1Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, China.

Abstract

OBJECTIVE:

To identify human umbilical cord mesenchymal stem cells (hUCMSCs) and increase the stability of the in vitro synthesized EGFP mRNA through modification.

METHODS:

Immunophenotype of hUCMSCs was examined by flow cytometry. Adipogenic and osteogenic differentiations were determined by oil red O and alkaline phosphatase staining, respectively. In vitro synthesized EGFP mRNA was modified through polyA tailing, capping and adding base analogues, and then transfected into the hUCMSCs. After transfection, the fluorescence intensity was detected by flow cytometry and cell viability was determined by Trypan blue staining.

RESULTS:

Flow cytometry revealed that the hUCMSCs were positive for CD29, CD44, CD105, and were negative for CD34, CD45 and HLA-DR. They had the capacity of differentiating into adipocytes and osteoblasts. Through the modification the in vitro synthesized mRNA was more stable than unmodified mRNA, and mRNA transfection had lower cytotoxicity than DNA.

CONCLUSION:

Modification can highly improve the stability of in vitro synthesized mRNA, which can be translated into protein in hUCMSCs.

PMID:
24103272
[PubMed - indexed for MEDLINE]
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