Cryopreservation and Laser Nanowarming of Zebrafish Embryos Followed by Hatching and Spawning

Overview

Journal Adv Biosyst

Specialties Biology
Biomedical Engineering
Biotechnology

Date 2020 Sep 30

PMID 32996298

Citations 15

Authors

Kanav Khosla

Joseph Kangas

Yilin Liu

Li Zhan

Jonathan Daly

Mary Hagedorn

John Bischof

Affiliations

Soon will be listed here.

Abstract

This study shows for the first time the ability to rewarm cryopreserved zebrafish embryos that grow into adult fish capable of breeding normally. The protocol employs a single injection of cryoprotective agents (CPAs) and gold nanorods (GNRs) into the yolk and immersion in a precooling bath to dehydrate the perivitelline space. Then embryos are encapsulated within CPA and GNR droplets, plunged into liquid nitrogen, cryogenically stabilized, and rewarmed by a laser pulse. Postlaser nanowarming, embryos (n = 282) exhibit intact structure by 1 h (40%), continued development after 3 h (22%), movement after 24 h (11%), hatching after 48 h (9%), and swimming after Day 5 (3%). Finally, from fish that survives till Day 5, two larvae are grown to adulthood and spawned, yielding survival comparable to an unfrozen control. Future efforts will focus on improving the survival to adulthood and developing methods to cryopreserve large numbers of embryos for research, aquaculture, and biodiversity preservation.

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