title: [Effects of Leukemia Inhibitory Factor Combined with Basic Fibroblast Growth Factor on Self-maintenance and Self-renewal of Human Umbilical Cord Mesenchymal Stem Cells In Vitro].
abstract: OBJECTIVE
  To study the effects of LIF combined with bFGF on the proliferation, stemness and senescence of hUC-MSC.
  
  
  METHODS
  Experiments were divided into 4 groups: control group, in which the cells were treated with complete medium (α-MEM containing 10% FBS); group LIF, in which the cells were treated with complete medium containing 10 ng/ml LIF; group bFGF, in which the cells were treated with complete medium containing 10 ng/ml bFGF; combination group, in which the cells were treated with complete medium containing 10 ng/ml LIF and 10 ng/ml bFGF. The growth curves of hUC-MSC at passage 4 in different groups were assayed by cell counting kit 8. Cellular morphologic changes were observed under inverted phase contrast microscope; hUC-MSC senescence in different groups was detected by β-galactosidase staining. The expression of PCNA, P16, P21, P53, OCT4 and NANOG genes was detected by RT-PCR.
  
  
  RESULTS
  The cell growth curves of each group were similar to the S-shape; the cell proliferation rate from high to low as follows: that in the combination group > group bFGF > group LIF > control group. Senescence and declining of proliferation were observed at hUC-MSC very early in control group; the cells in group LIF maintained good cellular morphology at early stage, but cell proliferation was slow and late senescence was observed; a few cells in group bFGF presented signs of senescence, but with quick proliferation; the cells in combination group grew quickly and maintained cellular morphology of hUC-MSC for long time. The LIF and bFGF up-regulated the expression of PCNA, OCT4 and NANOG, while they down-regulated the expression of P16, P21, P53, and their combinative effects were more significant.
  
  
  CONCLUSIONS
  LIF combined with bFGF not only can promote the proliferation and maintenance of stemness of hUC-MSC, but also can delay the senescence of hUC-MSC.
authors.affiliation: Medicine Graduate School, Nanchang University, Nanchang 330006, Jiangxi Province, China.|Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China.|Basic Medical College, Jiujiang University, Jiujiang 332000, Jiangxi Province, China.
authors.forename: Wen-Long
authors.initials: WL
authors.lastname: Hu
authors.affiliation: Medicine Graduate School, Nanchang University, Nanchang 330006, Jiangxi Province, China.
authors.forename: Ping-Ping
authors.initials: PP
authors.lastname: Wu
authors.affiliation: Basic Medical College, Jiujiang University, Jiujiang 332000, Jiangxi Province, China.
authors.forename: Chang-Chang
authors.initials: CC
authors.lastname: Yin
authors.affiliation: Medicine Graduate School, Nanchang University, Nanchang 330006, Jiangxi Province, China.|Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China.
authors.forename: Jian-Ming
authors.initials: JM
authors.lastname: Shi
authors.affiliation: Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China. E-mail: yinming0791@aliyun.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Yin
mesh_terms: D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005801:Genes, Homeobox; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D009930:Organic Chemicals; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology
chemical_list: C493956:CDKN1A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D053605:Leukemia Inhibitory Factor; D050814:Octamer Transcription Factor-3; D009930:Organic Chemicals; C494173:POU5F1 protein, human; D018809:Proliferating Cell Nuclear Antigen; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C420642:alpha minimal essential medium; D016222:Fibroblast Growth Factor 2