title: [Enrichment and Biological Characteristics of Peripheral Blood-derived Mesenchymal Stem Cells in Rats].
abstract: OBJECTIVE
  To explore the effective method for enrichment of rat peripheral blood-derived mesenchymal stem cells(PBMSC) and study the cell biological characteristics.
  
  
  METHODS
  Peripheral mononuclear cells were isolated by density gradient centrifugation from blood of 4 week old rats after G-CSF mobilization. Thereafter, the fibroblast-like cells were acquired by plastic-adherent culture, and the proliferation curve was assayed. For analyzing surface markers of the second generation cultured isolated PBMSC, both flow cytometry(CD90, CD44, CD29, CD45, CD11b and CD79a) and immunocytochemical staining(CD73, CD105, CD34 and HLA-DR) methods were used. Furthermore, the differentiation capacities of PBMSC into osteocytes, chondrocytes and adipocytes were identified.
  
  
  RESULTS
  (1) The adherent cells displayed typical colony-forming unit fibroblast(CFU-F) growth pattern after 6-7 day of primary culture and reached 80% confluence after 21 days of culture. The passaged PBMSC possessed high proliferative capacity and spindle growth pattern and was able to grown into exponential phase next day with a doubling time of 39.2 h. (2) PBMSC expressed mesenchymal markers such as CD90, CD44, CD29, CD73 and CD105, but failed to expressed markers of CD45, CD11b, CD79a, CD34 and HLA-DR. (3) After 21 days of culture in osteogenic, chondrogenic and adipogenic differentiation media, calcifying nodules, intracellular glycosaminoglycans and lipid droplets could be found by alizarin red, alcian blue and oil red-O staining, respectively.
  
  
  CONCLUSIONS
  PBMSC can be enriched from rat peripheral blood with high purity and abundance by our methods. The growth and phenotypic characteristics of the isolated PBMSC are consistent with that of well-known MSC, and these cells possess the capability to multi-lineage mesoderm differentiation.
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Lin-Ying
authors.initials: LY
authors.lastname: Jia
authors.affiliation: Department of Human Anatomy, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Fang
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Long
authors.initials: L
authors.lastname: Chen
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Li-Mei
authors.initials: LM
authors.lastname: Yu
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China.
authors.forename: Jin-Wei
authors.initials: JW
authors.lastname: Liu
authors.affiliation: Key Laboratory of Cell Engineering of Guizhou Province, Zunyi Medical College and Its Affiliated Hospital, Zunyi 563099, Guizhou Province, China. E-mail: oceanzt@163.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
mesh_terms: D017667:Adipocytes; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes; D005434:Flow Cytometry; D059630:Mesenchymal Stem Cells; D010011:Osteocytes; D051381:Rats