Abstract
Research question
Is vitrification with microinjection of single seminiferous tubules an efficient cryopreservation
approach for limited testicular tissue?
Design
Testicular tissue from 10 patients with normal spermatogenesis were assigned to a
fresh control group or one of the following cryopreservation procedures: uncontrolled
slow freezing (USF) using either 1.5 or 2.1 M DMSO combined with sucrose and vitrification
with or without single seminiferous tubules microinjection.
Results
Single seminiferous tubules microinjected with cryoprotective agents (CPA) enhanced
the penetration of CPA compared with CPA-treated testicular tissue fragments. Microinjection
of seminiferous tubules (VLP) maintained tubule structural integrity and germ cell
numbers, and reduced spermatogonial apoptosis after cryopreservation compared with
vitrification without microinjection (apoptosis rate: VLP versus vitrification without
microinjection, P = 0.047; VLP versus USF, P= 0.049). Freezing of single seminiferous tubules using 0.25-ml straws and traditional
sperm freezing methods protected sperm retrieval and recovery rates, and the progressive motility
index.
Conclusions
Vitrification of single seminiferous tubule with microinjection of low CPA concentration
is an effective approach to testicular cryopreservation.
KEYWORDS
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Biography
Dr Li Zheng is Professor of Department of Andrology at Shanghai General Hospital. His main research interests are fertility preservation and molecular mechanism of male infertility. With over 30 years’ experience in reproductive biology, he developed new carriers for cryopreservation of sperm and evaluated its safety, and established a rare sperm cryopreservation protocol and applied it in clinical practice which gave birth to healthy offspring.
Key message
Vitrification using 1.05-M DMSO and 1.35-M ethylene glycol for 10 min and 2.1-M DMSO and 2.7-M ethylene glycol for 3 min with microinjection was an efficient alternative to uncontrolled slow freezing for human testicular tissue. This protocol will provide an unprecedented cryopreservation solution for limited human testicular tissue.
Article info
Publication history
Published online: July 03, 2021
Accepted:
June 28,
2021
Received in revised form:
June 8,
2021
Received:
March 30,
2021
Declaration: The authors report no financial or commercial conflicts of interestIdentification
Copyright
© 2021 Published by Elsevier Ltd on behalf of Reproductive Healthcare Ltd.
