Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2011, Vol. 33 ›› Issue (6): 675-678.doi: 10.3881/j.issn.1000-503X.2011.06.018

• Original Articles • Previous Articles     Next Articles

Virus Free Induction of Umbilical Cord Derived Mesenchymal Stem Cells into Islet-like Cells

LI Jing , ZHU Li, ZHAO Chun-hua   

  1. Center of Excellence in Tissue Engineering, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing, 100005, China
  • Received:2011-11-11 Revised:2012-01-04 Online:2011-12-20 Published:2011-12-20
  • Contact: ZHAO Chun-hua E-mail:zhaoch16@hotmail.com
  • Supported by:

    Supported by the National Key Scientific Program of China(2011CB964901)

Abstract: Objective To explore the feasibility of using a virus-free system in the induction of umbilical cord derived mesenchymal stem cells (UC-MSCs) into insulin-secreting cells. Methods MSCs were isolated from human umbilical cord and induced into insulin-secreting cells with a three-stage method. The mRNA expression levels of foxa2, sox17, pdx1, ngn3, pax4, insulin, and glut-2 were compared between induced and non-induced groups by RT-PCR in each stage. The distribution pattern of insulin and c-peptide were detected by immunofluorescence staining and observed by fluorescence microscopy. Insulin and c-peptide secretion and glucose responsiveness were detected by enzyme-linked immunosorbent assay (ELISA). ResultsTranscription factors foxa2, sox17, pdx1, ngn3, pax4, insulin, and glut-2 were expressed in the induced cells. The mRNA expression levels of foxa2 and sox17 were significantly higher in the induced group than those in non-induced group in the first stage (all P<0.05), pdx1, ngn3, and pax4 were significantly higher in the induced cells than those in non-induced cells in the second stage (all P<0.05), and insulin and glut-2 expressions were significantly up-regulated in the induced group at last stage (all P<0.05). Immunofluorescence staining showed that insulin and c-peptide were located in the cytoplasm of more than 90% of induced cells. ELISA showed that total intracellular insulin content of the induced cells contained up to (346.3-739±32.5-149)μU/ml, which was significantly higher than insulin in non-induced cells (17.69±1.46) μU/ml (P<0.01). C-peptide content of the induced cells measured up to (195.10±8.88) pmol/L/h (P<0.01), when exposed to 5.5 mmol/L glucose (P<0.01). When stimulated with 22 mmol/L glucose, the c-peptide content of the induced cells increased to (340.99±7.91)pmol/L/h (P<0.01). Conclusion The umbilical cord derived MSCs can be efficiently induced into insulin-secreting cells via a virus-free system.

Key words: umbilical cord, mesenchymal stem cell, insulin, transcription factor

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