title: Programming of human umbilical cord mesenchymal stem cells in vitro to promote pancreatic gene expression.
abstract: Human umbilical cord mesenchymal stem cells (HUMSCs) are candidates for tissue engineering and may potentially be used for transdifferentiation into pancreatic endocrine cells. The adenoviral vector is effective in transducing genes into stem cells that are refractory to gene delivery by non‑viral approaches. qPCR was used to detect the pancreatic endogenous gene expression of HUMSCs transfected by islet cell-specific transcription factors (TFs). In the present study, using adenoviruses, the mouse TFs, pancreatic and duodenal homeobox 1 (pdx1), V-maf musculoaponeurotic fibrosarcoma oncogene homolog A (mafa) and class B basic helix‑loop‑helix factor neurogenin 3 (ngn3), which are essential for pancreatic cell development, were introduced into HUMSCs to assess the expression of the pancreatic genes, glucagon, pdx1 and nk2 homeobox 2 (nkx2.2). When pdx1, mafa and ngn3 were cotransduced into HUMSCs, the expression of glucagon increased by 21‑fold at days 3 and 7 following transduction, while the endogenous pdx1 gene expression was increased by 15‑fold at day 3 and decreased by 70% at day 7. When mafa and ngn3 were cotransduced into HUMSCs, there was a 5‑fold increase in pdx1 gene expression at day 7, but no activation was observed at day 3. When mafa alone was introduced into HUMSCs, the pdx1 gene expression was elevated by 6‑fold at day 3 and decreased by 3‑fold at day 7. Transduction of ngn3 alone into HUMSCs induced nkx2.2 gene expression at day 3 but the expression levels were decreased at day 7. However, when pdx1 and ngn3 were cotransduced into HUMSCs, the expression levels of glucagon, pdx1 and nks2.2 were all lower than those observed with pdx1 or ngn3 transduction alone. These results suggested that the transduction of pdx1, mafa and ngn3 genes into HUMSCs induced the expression of the pancreatic genes, glucagon, pdx‑1 and nkx2.2, and that the expression was time dependent. In addition, different combinations of the TFs may demonstrate synergistic or antagonistic effects. This data may be beneficial for guiding future studies obtaining mature pancreatic endocrine cells from HUMSCs.
authors.affiliation: Department of Pediatrics, The Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong 515041, P.R. China.
authors.forename: Hongwu
authors.initials: H
authors.lastname: Wang
authors.forename: Yuping
authors.initials: Y
authors.lastname: Yang
authors.forename: Guyu
authors.initials: G
authors.lastname: Ho
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Lin
authors.forename: Weizhao
authors.initials: W
authors.lastname: Wu
authors.forename: Weizhong
authors.initials: W
authors.lastname: Li
authors.forename: Limin
authors.initials: L
authors.lastname: Lin
authors.forename: Xueyong
authors.initials: X
authors.lastname: Feng
authors.forename: Xia
authors.initials: X
authors.lastname: Huo
authors.forename: Jikai
authors.initials: J
authors.lastname: Jiang
authors.forename: Xiaoshan
authors.initials: X
authors.lastname: Liu
authors.forename: Tianhua
authors.initials: T
authors.lastname: Huang
authors.forename: Chiju
authors.initials: C
authors.lastname: Wei
authors.forename: Lian
authors.initials: L
authors.lastname: Ma
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D054338:Cell Transdifferentiation; D065150:Cellular Reprogramming; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000378:metabolism; D005934:Glucagon / Q000235:genetics / Q000378:metabolism; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051473:Maf Transcription Factors, Large / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D014470:Umbilical Cord / Q000166:cytology*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005934:Glucagon; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D051473:Maf Transcription Factors, Large; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C485160:Mafa protein, mouse; C000715631:NKX2-2 protein, human; C407737:Neurog3 protein, mouse; C000715627:Nkx2-2 protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein