| In this study, umbilical cord mesenchymal stem cells (UCMSCs) were isolated from umbilical cordtissues of Luxi cattle. Surface markers and biological characteristics of them were then investigated.The results were as following:1. The UCMSCs were isolated from umbilical cord tissues of Luxi cattle by the method of tissueculture and the collagenase digestion. The optimal composition of culture media was L-DMEM+10%FBS+2mM glutamine. The UCMSCs were subcultured to as many as passage32and werecryopreserved for58vials in all.2. The markers of UCMSCs were detected by immunofluorescence staining and RT-PCR assay.The results showed that UCMSCs expressed CD29, CD44, CD73, CD90and CD16, but not expressedCD34, CD45and BLA-DR.3. The influences of cryoconservation and resuscitation on viability were evaluated. The resultsshowed that cell viability after cryopreservation was lower than before, with the cell viability before andafter cryopreservation about90%.4. The growth curves of different passages all showed the cell growth underwent latent period,logarithmic phase and plateau phase. The population doubling time of P3is about30.64h, and those forP8, P15P25are about30.94h,31.80h and40.65h respectively. We can see, the population doublingtime is prolonged along with the increasing passage number and the ability of cell proliferation isdeclined.5. Colony forming cell assay of different passages showed that the cloning efficiency were (52.67±5.03)%,(43.67±9.30）%,(28.33±1.53)%,(4.00±2.65)%corresponding P3, P8, P15, P25respectively. In short, colony formation faded with increasing passage number.6Cell cycle progression of different passages showed that most of the generations of cells in theG0/G1phase, and the proportion of S phase cells reduced gradually with increasing passage.7The UCMSCs of Luxi cattle have multi-lineage differentiation potential. Upon proper induction,UCMSCs could differentiate into osteoblasts, adipocytes, liver-like cells, islet-like cells and nerve cells. |
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