title: In vitro differentiation of bone marrow mesenchymal stem cells into endometrial epithelial cells in mouse: a proteomic analysis.
abstract: OBJECTIVE
  Mouse bone marrow mesenchymal stem cells (BMSCs) have been demonstrated to differentiate into female endometrial epithelial cells (EECs) in vivo. Our previous studies demonstrated that BMSCs can differentiate in the direction of EECs when co-cultured with endometrial stromal cells in vitro. Here, we obtain and analyse differential proteins and their relevant pathways in the process of BMSCs differentiating into EECs by isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis.
  
  
  METHODS
  A 0.4-μm pore size indirect co-culture system was established with female mice endometrial stromal cells (EStCs) restricted in the upper Transwell chamber and BMSCs in the lower well plate. After indirect co-culture for several days, the BMSCs were revealed to progressively differentiate towards EECs in vitro. Then, four groups were divided according to different co-culture days with single culture groups of BMSCs as controls. Proteins were detected using iTRAQ based on 2DLC-ESI-MS/MS and data were analysed by bioinformatics.
  
  
  RESULTS
  A total number of 311 proteins were detected, of which 210 proteins were identified with relative quantitation. Among them, 107 proteins were differentially expressed with a 1.2-fold change as the benchmark, with 61 up-regulated and 46 down-regulated proteins. Differential proteins CK19 and CK8 were epithelial markers and upregulated. Stromal marker vimentin were downregulated. Top canonical pathways was "remodeling of epithelial adhesions junctions" and "actin cytoskeleton signaling". Top networks was "cell-to-cell signaling and interaction, tissue development and cellular movement" regulated by ERK/MAPK and α-catenin.
  
  
  CONCLUSIONS
  To the best of our knowledge, this is the first preliminary study of differential protein expression in the differentiation process of BMSCs into EECs in vitro. We further elucidated BMSCs differentiated in the direction of EECs. In addition, ERK/MAPK and α-catenin played important roles by regulating core differential proteins in the "cell-to-cell signaling and interaction, tissue development and cellular movement" network.
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Cong
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Yisheng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Wenbi
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Mingjun
authors.initials: M
authors.lastname: Cheng
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Obstetrics and Gynecology Hospital of Fudan University, Fudan University Shanghai, P.R. China ; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Fudan University Shanghai, P.R. China ; Department of Obstetrics and Gynecology of Shanghai Medical School, Fudan University Shanghai, P.R. China ; Institute of Biomedical Sciences, Fudan University Shanghai, P.R. China.
authors.forename: Congjian
authors.initials: C
authors.lastname: Xu
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D018920:Coculture Techniques; D004717:Endometrium / Q000166:cytology* / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
keywords: Bone marrow mesenchymal stem cells; differentiation; endometrial epithelial cells; proteomic analysis