title: DNA methylation patterns of imprinting centers for H19, SNRPN, and KCNQ1OT1 in single-cell clones of human amniotic fluid mesenchymal stem cell.
abstract: OBJECTIVE
  To test the hypothesis that human amniotic fluid mesenchymal stem cells contain a unique epigenetic signature in imprinting centers of H19, SNRPN, and KCNQ1OT1 during in vitro cell culture.
  
  
  METHODS
  By bisulfite genomic sequencing, we analyzed the imprinting centers of three imprinted genes (including H19, SNRPN, and KCNQ1OT/) in a total of six single-cell clones of human amniotic fluid mesenchymal stem cells at cell passages 7, 8, 9, and 10 during in vitro cell culture.
  
  
  RESULTS
  The imprinting centers of H19 and KCNQ1OT1 showed hypermethylation at passage 7 in all single-cell clones of human amniotic fluid mesenchymal stem cells, and there was no significant change in DNA methylation patterns during in vitro cell culture. The imprinting centers of SNRPN showed variable methylation patterns at passage 7 in six single-cell clones, and DNA methylation patterns varied during in vitro cell culture from passages 8 to 10.
  
  
  CONCLUSIONS
  In conclusion, human amniotic fluid mesenchymal stem cells contain a unique epigenetic signature during in vitro cell culture. H19 and KCNQ1OT1 possessed a substantial degree of hypermethylation status, and variable DNA methylation patterns of SNRPN was observed during in vitro cell culture of human amniotic fluid mesenchymal stem cells. Our results urge further understanding of epigenetic status of human amniotic fluid mesenchymal stem cells before it is applied in cell replacement therapy.
authors.affiliation: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Chang Gung University, Tao-Yuan, Taiwan.
authors.forename: Hsiu-Huei
authors.initials: HH
authors.lastname: Peng
authors.forename: Shuenn-Dyh
authors.initials: SD
authors.lastname: Chang
authors.forename: An-Shine
authors.initials: AS
authors.lastname: Chao
authors.forename: Chao-Nin
authors.initials: CN
authors.lastname: Wang
authors.forename: Po-Jen
authors.initials: PJ
authors.lastname: Cheng
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.forename: Tzu-Hao
authors.initials: TH
authors.lastname: Wang
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D002999:Clone Cells; D019175:DNA Methylation; D018392:Genomic Imprinting; D006801:Humans; D059630:Mesenchymal Stem Cells; D016133:Polymerase Chain Reaction; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics; D062085:RNA, Long Noncoding / Q000235:genetics*; D017422:Sequence Analysis, DNA; D055517:snRNP Core Proteins / Q000235:genetics*
chemical_list: C094335:H19 long non-coding RNA; C485621:KCNQ1OT1 long non-coding RNA, human; D024642:Potassium Channels, Voltage-Gated; D062085:RNA, Long Noncoding; C526417:SNRPN protein, human; D055517:snRNP Core Proteins