Proliferation capacity of rabbit adipose tissues-derived mesenchymal stem cells at early stage in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND:Surface marker molecules and differential ability of human adipose tissues-derived mesenchymal stem cells can change with the prolonged culture time.OBJECTIVE:To analyze the morphological characteristics and colony forming capacity of early cultured rabbit adipose tissues-derived mesenchymal stem cells in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Ophthalmology,and Central Laboratory,Eye and ENT Hospital,Fudan University from January to March 2008.MATERIALSNine female New Zealand rabbits aged three months were obtained from the Shanghai Yingenyang Rabbit Room.METHODS:Adipose tissue was harvested from the nape fat pad of the New Zealand rabbits following anesthesia.Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 0.1 volume fraction of fetal bovine serum.Cells were passaged when 80% was confluent.The second,third and fourth passages of cells were utilized for the study.MAIN OUTCOME MEASURES:The morphology of adipose tissues-derived mesenchymal stem cells was observed using inverted microscope.Cell phenotype was detected utilizing flow cytometry.The cloning capacity was determined.RESULTS:Adipose tissues-derived mesenchymal stem cells were fibroblast-like and proliferated rapidly in vitro at the second,third and fourth passage.Flow cytometry showed that adipose tissues-derived mesenchymal stem cells expressed CD29 and PCNA.The colony-forming rates were respectively(8.0?0.6)%,(6.7?0.4)% and(4.6?0.5)%(F=12.18,P