Vitreous cryopreservation of tenocytes co-cultured with porous polydimethylsiloxane scaffolds / 中国组织工程研究

ABSTRACT

BACKGROUND:Accumulative evidence supports that vitreous cryopreservation can improve the cel survival rate. OBJECTIVE:To investigate the effect of vitreous cryopreservation on the tenocytes co-cultured with the porous polydimethylsiloxane (PDMS) scaffold. METHODS:Tenocytes were co-cultured with the porous PDMS scaffold for 9-14 days, and then preserved and resuscitated in the 10%dimethyl sulfoxide (DMSO), 21%DMSO and VS55, respectively. One hour later, the survival rate of post-resuscitated tenocytes versus pre-resusciated tenocytes was analyzed by live/dead double color fluorescent staining and flow cytometry. RESULTS AND CONCLUSION:Live/dead double color fluorescent staining revealed that tenocytes in the 10%DMSO group appeared to be irregular and double stained, and a large number of cel s shedding from the scaffold. The VS55 and 21%DMSO groups showed some spindle and hemispherical cel s single stained for green fluorescence and few double stained irregular cel s. Additional y, the cel density in the two groups was significantly lower than that in the control group. Flow cytometry results found that there were homogenous cel s in the control group;the number of cel s in the 10%DMSO group was too low to undergo flow cytometry;smal cel particles were visible in the VS55 group;in the 21%DMSO group, the cel volume was similar with the control group, and smal particles also existed. The survival rate in the VS55 group (64.9%) was significantly lower than that in the 21%DMSO group (76.2%;P<0.05). Conversely, the survived cel s were rare in the 10%DMSO group. To conclude, 21%DMSO vitreous cryopreservation improves the cel survival rate and is beneficial for tenocyte adherence to the scaffold.