Du T.-Q.,Shandong University |
Chi L.-L.,Shandong Qilu Stem Cells Co. |
Zhao S.-Q.,Zaoihuang Maternal and Child Care Service Hospital |
Shi Q.,Shandong University |
And 4 more authors.
Acta Anatomica Sinica | Year: 2015
Objective To study the cffects of freezing sheep intact ovary by perfusing different cryoprotectants with concentration gradient blended programmed organ cooling perfusor, and to obtain the best cryoprotectants combination for cryopreservation of the whole sheep ovary. Methods Twenty-eight ovaries collected from 6-8 months non-pregnant female sheeps were randomly distributed into fresh group (group A) , trehalose group (group B) , glycerin group (group C) and DMSO group (group D). The morphology, cell apoptosis (by HE staining and TUNEL assay) and mRNA transcript of Bcl-2 associated X protein and cold inducible RNA-binding protein (by real-time PCR) of thawed sheep ovaries were tested to established the criterion for appraising cryopreservation results. Results The percentage of normal follicles in group B was comparable with group A (P > 0. 05). The value in group C and group D were significant lower than that in group A (P<0. 05). Quantitative assessment of stromal cell density indicated that the values in group D significantly lower than values in group A while group B and group C had values similar to those of group A. The TUNEL assay showed that the number of positive cells in group A were the lowest of all groups followed by group B, group C and group D (P < 0. 05). The level of BAX transcripts significantly increased in group C and group D (P < 0. 05). The level of CIRP transcripts increased highest in group B followed by group C and group D (P < 0. 05). Conclusion The whole ovary can be appropriately preserved after cryopreservation, and cryoprotectant combination of group B appears to be better for cryopreservation of whole sheep ovary in aspect of histology and anti-apoptosis.
Du T.,Nanjing Medical University |
Chao L.,Shandong University |
Zhao S.,Zaozhuang Maternal and Children Health Hospital |
Chi L.,Shandong Qilu Stem Cells Co. |
And 4 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2015
Purpose: The study aims to assess the protective effects of dimethyl sulfoxide (DMSO)-free solution based on trehalose on the cryopreservation of a whole sheep ovary and evaluate its use as an efficient cryoprotectant. Method: Twenty-one ovaries collected from 6- to 8-month-old non-pregnant female sheep were randomly distributed into three groups, namely, a fresh group, a DMSO-free group, and a DMSO group. The morphology, cell apoptosis (by hematoxylin and eosin (HE) staining and terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay), and mRNA transcript of Bcl-2-associated X protein (BAX) and cold inducible RNA-binding protein (CIRP) (by real-time PCR) of the thawed sheep ovaries and fresh controls were tested to establish a criterion for appraising the results of the cryopreservation. Results: (i) The histological assessment indicated that the structure of the DMSO-free ovaries remained largely intact and comparable to those of the fresh control groups; whereas, significant damage was observed in the ovaries of the DMSO group (P < 0.05). (ii) The TUNEL assay and mRNA transcript of the BAX assessment showed that the apoptosis parameter in the fresh group was the lowest among all the groups (P < 0.05), and the parameter in the DMSO-free group was significantly lower than that in the DMSO group (P < 0.05). (iii) The level of the CIRP transcripts increased the most in the DMSO-free group followed by the DMSO group and the fresh control group (P < 0.05). Conclusions: These results indicate that a DMSO-free cryoprotectant solution, especially a trehalose cryoprotectant, is an efficient cryoprotectant and has a beneficial effect on the cryopreservation of whole sheep ovaries. © 2015, Springer Science+Business Media New York.
Wang D.-K.,Shandong University |
Wang Z.-H.,Shandong Qilu Stem Cells Co. |
Guo H.,Shandong Qilu Stem Cells Co. |
Zhang L.-L.,Shandong University
Chinese Journal of Tissue Engineering Research | Year: 2013
BACKGROUND: General assessment of viability of umbilical cord mesenchymal stem cells during transport is not considered at home and abroad. OBJECTIVE: To explore the effects of different transportation conditions such as albumin and transport time on survival rate of umbilical cord mesenchymal stem cells. METHODS: Umbilical cord mesenchymal stem cells cultured in vitro were divided into experimental and control groups. Each of group contained 3.15×109/L cells in 3 mL normal saline. The experimental group contained 1% albumin in dark environment. The control group included two subgroups: dark preservation group with the absence of albumin and light preservation group with the presence of albumin. Cell counting, trypan blue staining and cell survival rate were compared at 0, 2, 4, 8, and 24 hours. RESULTS AND CONCLUSION: Compared with the control group, no cell loss was observed in experimental group (with the presence of albumin in the dark) and a 90% viability ratio was achieved at 8 hours. In the controlgroup without albumin, loss rate reached 38.5% and survival rate reached 86% at 8 hours. Results revealed that 1% albumin predominantly improved cell survival rate in long-distance transport of umbilical cord mesenchymal stem cells. When transport time was more than 8 hours, cell loss rate increased and cell survival rate downregulated. Our experimental data demonstrated that umbilical cord mesenchymal stem cells preserved in normal saline consisting of 1% albumin placing in dark environment at 16 °C apply to clinical application criterion in 8 hours.