Open Access

miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells

  • Anahita Shaer1, 2,
  • Negar Azarpira2, 4Email author,
  • Akbar Vahdati1, 2,
  • Mohammad Hosein Karimi2 and
  • Mehrdad Shariati3
Cellular & Molecular Biology LettersAn International Journal201419:207

https://doi.org/10.2478/s11658-014-0207-3

Received: 10 November 2013

Accepted: 19 August 2014

Published: 29 August 2014

Abstract

This paper focuses on the development of renewable sources of isletreplacement tissue for the treatment of type I diabetes mellitus. Placental tissue-derived mesenchymal stem cells (MSCs) are a promising source for regenerative medicine due to their plasticity and easy availability. They have the potential to differentiate into insulin-producing cells. miR-375 is a micro RNA that is expressed in the pancreas and involved in islet development. Human placental decidua basalis MSCs (PDB-MSCs) were cultured from full-term human placenta. The immunophenotype of the isolated cells was checked for CD90, CD105, CD44, CD133 and CD34 markers. The MSCs (P3) were chemically transfected with hsa-miR-375. Total RNA was extracted 4 and 6 days after transfection. The expressions of insulin, NGN3, GLUT2, PAX4, PAX6, KIR6.2, NKX6.1, PDX1, and glucagon genes were evaluated using real-time qPCR. On day 6, we tested the potency of the clusters in response to the high glucose challenge and assessed the presence of insulin and NGN3 proteins via immunocytochemistry. Flow cytometry analysis confirmed that more than 90% of the cells were positive for CD90, CD105 and CD44 and negative for CD133 and CD34. Morphological changes were followed from day 2. Cell clusters formed during day 6. Insulin-producing clusters showed a deep red color with DTZ. The expression of pancreatic-specific transcription factors increased remarkably during the four days after transfection and significantly increased on day 7. The clusters were positive for insulin and NGN3 proteins, and C-peptide and insulin secretion increased in response to changes in the glucose concentration (2.8 mM and 16.7 mM). In conclusion, the MSCs could be programmed into functional insulin-producing cells by transfection of miR-375.

Keywords

Pancreas Beta cells miR-375 Placenta Mesenchymal stromal cells Induced pluripotent stem cell microRNA Insulin Differentiation NGN3 GLUT2 PDX1

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