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Clinical and Experimental Medicine

, Volume 15, Issue 4, pp 501–509 | Cite as

PDX-1 mRNA-induced reprogramming of mouse pancreas-derived mesenchymal stem cells into insulin-producing cells in vitro

  • Xing Rong Guo
  • Xiao Li Wang
  • Man Chol Li
  • Ya Hong Yuan
  • Yun Chen
  • Dan Dan Zou
  • Liu Jiao BianEmail author
  • Dong Sheng LiEmail author
Original Article

Abstract

Pancreatic islet transplantation has remained an effective therapy for type 1 diabetes since 2000. Its widespread use has been prohibited by the shortage of suitable donors. It is critical to explore an applicable alternative for β-cell replacement. This study was performed to generate insulin-producing cells (IPCs) from pancreas-derived mesenchymal stem cells (pMSCs). pMSCs were isolated from discarded pancreatic tissue in the filter liquor during islet isolation procedure in mice and ex vivo expanded in culture. IPCs were induced by transfection of pancreas and duodenal transcription factor 1 (PDX-1) mRNA in vitro. Some islet characteristics were identified on pMSC-derived IPCs in mRNA and protein levels. Our results demonstrated that mouse pMSCs can be transdifferentiated into effective glucose-responsive insulin-producing cells through transfecting synthetic modified PDX-1 mRNA in vitro. The study of PDX-1 mRNA-induced pMSC reprogramming may pave the way toward the development of a novel β-cell source for the treatment of diabetes.

Keywords

Pancreas-derived mesenchymal stem cells (pMSCs) Reprogramming PDX-1 mRNA Insulin-producing cells (IPCs) 

Notes

Acknowledgments

This study was financially supported by the Major Science and Technology Projects of China (No. 2013ZX10001-004-002-005), National Natural Science Foundation of China (No. 21075097), the National Natural Science Foundation of China (81070614), National Natural Science Foundation of Hubei province (2012FFA037), Natural Science Support Foundation for graduated students of Hubei University of Medicine (2011QDZR-8) and Foundation of Hubei Educational Committee(B20122411). Authors are grateful to Dr. Long-Jun Dai of University of British Columbia in Canada for his critical review and discussion of the manuscript.

Conflict of interest

None.

Ethical standards

All animal studies have been approved by the appropriate ethics committee and have therefore been performed in accordance with the ethical standards.

Supplementary material

10238_2014_319_MOESM1_ESM.tif (1.8 mb)
Supplementary Material Figure 1. A. Immunofluorescence analyses of PDX-1 expression in islets and islet-derived MSCs. Fresh islets cultured for 8 days in modified RPMI 1640 media containing 20 ng/ml bFGF and EGF showing outgrowth of the monolayer of cells—islet-derived MSCs. PDX-1 specially expressed in islets, but undetected in islet-derived MSCs, which is consistent with the pMSCs obtained from our method. (black↑ for Islets, red↑ for pMSCs) B. GFP mRNA transfected into pMSCs as a control. The expression of GFP was determined with FACS (a) and examined under a fluorescent microscope (b). C. Morphological shape of differentiated pMSCs into IPCs in the first (a) and second approaches (b) on 8 days. (TIFF 1839 kb)
10238_2014_319_MOESM2_ESM.tif (2.2 mb)
Supplementary Material Figure 2. The primary culture of the isolated cells contained a heterogeneous cell population with both round and fibroblast-like cells. Round cells grew fast and fibroblast-like cells hardly grew on day 3(A); fibroblast-like cells began to grow, round cells stop developing on day 6(B); fibroblast-like cells proliferated very well on day 10(C). (black↑for round cells, red↑for pMSCs) (TIFF 2298 kb)
10238_2014_319_MOESM3_ESM.pdf (123 kb)
Supplementary Material Table 1. Primer sequences and PCR product size (PDF 122 kb)

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Copyright information

© Springer-Verlag Italia 2014

Authors and Affiliations

  • Xing Rong Guo
    • 1
    • 2
  • Xiao Li Wang
    • 2
  • Man Chol Li
    • 2
  • Ya Hong Yuan
    • 2
  • Yun Chen
    • 2
  • Dan Dan Zou
    • 2
  • Liu Jiao Bian
    • 1
    Email author
  • Dong Sheng Li
    • 2
    Email author
  1. 1.College of Life SciencesNorthwest UniversityXi’anChina
  2. 2.Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe HospitalHubei University of MedicineShiyanChina

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