P-218 Mitochondria transfer from adipose stem cell ameliorates the development potential of cryopreserved oocytes
Authors: Kankanam Gamage S.U , Hashimoto S , Miyamoto Y , Nakano T , Yamanaka M , Koike A , Satoh M , Morimoto Y 
Published: June 2022

Abstract

Study question
Could autologous adipose stem cell (ASC) mitochondria supplementation restore the post-fertilization development potentialin vitrified-thawed oocytes?

Summary answer
Autologous ASC mitochondria supplementation could rejuvenate the quality of vitrified-thawed oocytes and enhance the embryo’s developmental capacity.

What is known already
Pre- and post-implantation developmental potential of embryos derived from vitrified oocytes are remarkedly lower than those of fresh oocytes. Despite its’ low efficacy, oocyte cryopreservation is becoming prevalent in assisted reproductive technologies to cater the growing demands due to patients' sociological and pathological conditions. Unfavorable effects to mitochondrial membrane potential, mitochondrial structure, ATP production, reactive oxygen species (ROS), meiotic spindle and microfilaments, and Ca2+ ion regulation are reported as detrimental damages in oocytes following the cryopreservation, that adversely affect the development potential. Furthermore, autologous stem cell mitochondria supplementation can rescue the aging-related oocyte mitochondrial damages.

Study design, size, duration
The mature oocytes, autologous ASC, and mitochondria were collected from young mice and analyzed at Osaka City University, Japan. In total, 600 young mouse mature oocytes were occupied in this prospective study.

Participants/materials, setting, methods
Young C57BL/6JJmsmouse (8weeks) mature oocytes that have been pre-vitrified were thawed before the start of the ICSI procedure.ASC specificity and ASC mitochondria function and ultrastructure were pre-analyzed. ASC mitochondria were isolated on the same day and supplemented with intracellular sperm injection (ICSI) and as the control, the mitochondrial buffer was injected. The survival rate, fertilization rate, blastulation, mitochondria function, ROS level in 2 cell embryos, and live birth rates were compared between the 2 groups.

Main results and the role of chance
The ASC mitochondriashowed higher membrane potential compared to the somatic cells and were spherical in shape with low cristae numbers. The survival rate and the fertilization ratewere comparable in both mitochondria supplemented and control groups. However, theASC mitochondria supplementation seemed to havesignificantly improvedthe blastocyst development capacity from 2cell embryos compared to the control group (P &lt; 0.05;56.8% & 38.2%, respectively).And interestingly, a significantly higher ATP level was found in the mitochondria supplemented group’s 2 cell embryos thanin the control group (P &lt; 0.05;905.6pmol & 561.1pmol respectively). And though it was not statistically significant, a higher potential ofgetting live birth was found in the mitochondria supplemented group thanthe control group after 2 cell embryo transfer.

Limitations, reasons for caution
We acknowledge that the absence of compared data with fresh oocytes’ ICSI, the detailed cellular mechanism behind theimprovement of embryo development, and transgenerational safety in offspring developed fromthe mitochondria supplementation werethe limitations of this study.

Wider implications of the findings
With these results, we propose that ASC mitochondria supplementation could rejuvenate the quality of cryopreserved oocytes and enhance the embryo developmental capacity, signifying another possible approach of mitochondrial transplantation therapy.

Trial registration number
not applicable