title: Suspension Culture Alters Insulin Secretion in Induced Human Umbilical Cord Matrix-Derived Mesenchymal Cells.
abstract: OBJECTIVE
  Worldwide, diabetes mellitus (DM) is an ever-increasing metabolic disorder. A promising approach to the treatment of DM is the implantation of insulin producing cells (IPC) that have been derived from various stem cells. Culture conditions play a pivotal role in the quality and quantity of the differentiated cells. In this experimental study, we have applied various culture conditions to differentiate human umbilical cord matrix-derived mesenchymal cells (hUCMs) into IPCs and measured insulin production.
  
  
  METHODS
  In this experimental study, we exposed hUCMs cells to pancreatic medium and differentiated them into IPCs in monolayer and suspension cultures. Pancreatic medium consisted of serum-free Dulbecco's modified eagle's medium Nutrient mixture F12 (DMEM/F12) medium with 17.5 mM glucose supplemented by 10 mM nicotinamide, 10 nM exendin-4, 10 nM pentagastrin, 100 pM hepatocyte growth factor, and B-27 serum-free supplement. After differentiation, insulin content was analyzed by gene expression, immunocytochemistry (IHC) and the chemiluminesence immunoassay (CLIA).
  
  
  RESULTS
  Reverse transcription-polymerase chain reaction (RT-PCR) showed efficient expressions of NKX2.2, PDX1 and INSULIN genes in both groups. IHC analysis showed higher expression of insulin protein in the hanging drop group, and CLIA revealed a significant higher insulin production in hanging drops compared with the monolayer group following the glucose challenge test.
  
  
  CONCLUSIONS
  We showed by this novel, simple technique that the suspension culture played an important role in differentiation of hUCMs into IPC. This culture was more efficient than the conventional culture method commonly used in IPC differentiation and cultivation.
authors.affiliation: Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Seyedi
authors.affiliation: Stem Cell Research Lab, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Farsinejad
authors.affiliation: Department of Laboratory Medicine and Center for Bioscience, Karolinska Institute, Huddinge, Sweden.
authors.forename: Seyed Amirmahdi
authors.initials: SA
authors.lastname: Nematollahi-Mahani
authors.affiliation: Pharmaceutics Research Center (PRC), Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran.
authors.forename: Touba
authors.initials: T
authors.lastname: Eslaminejad
authors.affiliation: Department of Anatomy, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran; Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran; Afzal Research Institute, Kerman, Iran.
authors.forename: Seyed Noureddin
authors.initials: SN
authors.lastname: Nematollahi-Mahani
keywords: Insulin Producing Cell; Suspension Culture; Wharton Jelly Cells