title: Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells.
abstract: BACKGROUND
  Diabetes mellitus (DM) is an incurable metabolic disease constituting a major threat to human health. Insulin-producing cells (IPCs) differentiated from mesenchymal stem cells (MSCs) hold great promise in the treatment of DM. The development of an efficient IPC induction system is a crucial step for the clinical application of IPCs for DM. Laminin 411 is a key component of the basement membrane and is involved in the regulation of cell differentiation; however, little is known about a role of laminin 411 in the regulation of IPC differentiation from human MSCs.
  
  
  METHODS
  MSCs were isolated from human umbilical cord (UC-MSCs) and expanded in an in vitro culture system. UC-MSCs were then cultured in the IPC induction and differentiation medium in the presence of laminin 411. Flow cytometry, Quantitative realtime PCR, immunofluorescence staining, ELISA, Western blotting and other techniques were applied to determine IPC generation, insulin expression and related mechanisms. To evaluate potential therapeutic efficacy of IPCs induced from UC-MSCs, a type-1 diabetes (T1DM) rat model was generated using streptozotocin. Blood glucose, insulin levels, and survival of rats were monitored periodically following intravenous injection of the tested cells.
  
  
  RESULTS
  Laminin 411 markedly induced the expression of the genes Foxa2 and Sox17, markers for pancreatic precursor cells, efficiently induced IPC differentiation from MSCs, and up-regulated insulin expression at both mRNA and protein levels. Furthermore, the expression of the genes known to govern insulin expression including Pdx1 and Ngn3 was markedly induced by laminin 411, which suggests that Pdx1 and Ngn3 signaling pathways are involved in laminin 411 induced-insulin expression machinery. More importantly, administration of laminin 411-induced IPCs rapidly and significantly down-regulated fasting blood glucose levels, significantly reduced the HbA1c concentration and markedly improved the symptoms and survival of T1DM rats.
  
  
  CONCLUSIONS
  Our results demonstrate that laminin 411 acts as a potent differentiation inducer of IPCs from UC-MSCs via the Pdx1 and Ngn3 signaling pathways. Moreover, transfusion of laminin 411 induced-IPCs more efficiently improves symptoms and survival of T1DM rats. These novel finding highlights a potential clinical application of laminin 411 induced-IPCs in the treatment of T1DM, which calls for further studies.
authors.forename: Huiting
authors.initials: H
authors.lastname: Qu
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liu
authors.forename: Yihong
authors.initials: Y
authors.lastname: Ni
authors.forename: Yang
authors.initials: Y
authors.lastname: Jiang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Feng
authors.forename: Juan
authors.initials: J
authors.lastname: Xiao
authors.forename: Yanan
authors.initials: Y
authors.lastname: Guo
authors.forename: Dexiao
authors.initials: D
authors.lastname: Kong
authors.forename: Ai
authors.initials: A
authors.lastname: Li
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Li
authors.forename: Xianghua
authors.initials: X
authors.lastname: Zhuang
authors.forename: Zhilun
authors.initials: Z
authors.lastname: Wang
authors.forename: Yongjing
authors.initials: Y
authors.lastname: Wang
authors.forename: Yali
authors.initials: Y
authors.lastname: Chang
authors.forename: Shihong
authors.initials: S
authors.lastname: Chen
authors.forename: Feng
authors.initials: F
authors.lastname: Kong
authors.forename: Xuhua
authors.initials: X
authors.lastname: Zhang
authors.forename: Shengtian
authors.initials: S
authors.lastname: Zhao
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.forename: Dawei
authors.initials: D
authors.lastname: Xu
authors.affiliation: Departments of Hematology and Cellular Therapy, the Second Hospital of Shandong University, Jinan, Shandong, PR China. wdq5445@163.com.
authors.forename: Daoqing
authors.initials: D
authors.lastname: Wang
authors.forename: Chengyun
authors.initials: C
authors.lastname: Zheng
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000502:physiology*; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000096:biosynthesis*; D007797:Laminin / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D017931:DNA Primers; D007328:Insulin; D007797:Laminin