title: [Differentiation of mesenchymal stem cells induced by co-culture method].
abstract: OBJECTIVE
  To investigate the potential differentiation of human mesenchymal stem cells(MSCs)into epithelium-like cells by an in vitro co-culture method.
  
  
  METHODS
  The human conjunctival epithelium was obtained by digestion with dispase2, and the MSCs were isolated by density gradient centrifugalization. All cells were identified according to their morphologies and cell-surface antigen profiles by immunocytochemical analysis. The MSCs underwent co-culture with conjunctival epithelium in the manner without cell-to-cell contract. The morphological characterizes of cells were observed under contrast microscope, and the cytokeratin-4 expressions of the differentiated cells were identified by immunocytochemistry staining, reverse transcriptase polymerase chain reaction(RT-PCR), and Western blotting.
  
  
  RESULTS
  Immunocytochemistry showed that positive expression of CD29 and negative expression of CD34 in the in vitro cultured MSCs. Cytokeratins4(CK4)was positively expressed in the human conjunctival epithelium. After co-cultured with conjunctival epithelial for 10 days, CK4 was detected in differentiated cells by immunocytochemistry, RT-PCR, and Western blotting.
  
  
  CONCLUSIONS
  MSCs can differentiate into epithe1ium-like cells after having been co-cultured with human conjunctival epithelium.
authors.affiliation: Department of Ophthalmology, the Second Hospital of Shandong University, Jinan 250033, China.
authors.forename: Guo-ling
authors.initials: GL
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, the Second Hospital of Shandong University, Jinan 250033, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Ophthalmology, the Second Hospital of Shandong University, Jinan 250033, China.
authors.forename: Yang-yang
authors.initials: YY
authors.lastname: Du
authors.affiliation: Department of Anatomy, School of Medicine, Shandong University, Jinan 250012, China.
authors.forename: Yan-li
authors.initials: YL
authors.lastname: Liu
authors.affiliation: Department of Anatomy, School of Medicine, Shandong University, Jinan 250012, China.
authors.forename: Zhi-yu
authors.initials: ZY
authors.lastname: Liu
authors.affiliation: Department of Ophthalmology, the Second Hospital of Shandong University, Jinan 250033, China.
authors.forename: Han
authors.initials: H
authors.lastname: Zhang
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction