Protection of reduced glutathione and ulinastatin on frozen-thawed ovarian tissues after xenotransplantation

doi:10.3969/j.issn.2095-4344.1074

Abstract

Abstract:

BACKGROUND: Reduced glutathione, as an effective antioxidant can inhibit inflammatory cytokines. Ulinastatin can directly remove oxygen free radicals and inhibit production of oxygen free radicals and various inflammatory mediators, sustain the stability of cell membrane and lysosomal membrane, and inhibit the activity of active enzymes, thus protecting the transplanted organ.
OBJECTIVE: To evaluate the effect of reduced glutathione and ulinastatin on improving the xenotransplantation efficiency of frozen-thawed human ovarian tissue.
METHODS: Ovarian tissue samples from 6 women aged 31.8±5.7 years old in the First Affiliated Hospital, Sun Yat-sen University, were collected and each tissue sample was randomly divided into four groups after slow freezing: reduced glutathione, ulinastatin, glutathione + ulinastatin and blank control (normal saline) groups. Ovarian tissues were xenotransplantated under the back skin of male NOD-SCID mice after 15-minute cultivation. The mice were administered with corresponding drugs for 5 consecutive days. The grafts were collected at 1, 3, 7, 14, 28, 56, and 85 days after transplantation. Histomorphology, growth situation and viability of the follicle were observed through hematoxylin-eosin staining and enzymatic isolation method.
RESULTS AND CONCLUSION: The majority of the frozen-thawed ovarian tissues were primordial follicles (79.5±14.3%). The viability of the follicle in fresh ovarian tissue was 97.6%, and (84.4±5.7)% in thawed ovarian tissue. The reduced glutathione, ulinastatin, and glutathione + ulinastatin groups showed significantl improvement in the viability of the follicle compared with the control group (P=0.022). There was no significant difference between single and combined drug groups (P > 0.05). The follicle viability reached a lowest value at 3 days after transplantation (P < 0.05), and increased after 1 week later. The mean survival rate of the follicle in the isolated follicle at different time points showed no significant difference among groups (P=0.114). Compared with the histological sections, the isolated follicle showed highest survival rate. Our results suggest that reduced glutathione and ulinastatin can increase the early viability of the follicles after xenotransplantation, and combined administration shows no extra benefit.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Ovary, Cryopreservation, Glutathione, Transplantation, Tissue Engineering

CLC Number:

R446
R318

Long Huidong, Long Lingli, Mai Qingyun, Zhao Wen, Li Yubin. Protection of reduced glutathione and ulinastatin on frozen-thawed ovarian tissues after xenotransplantation[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(7): 1084-1089.

Figures/Tables 4

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