Offspring production from cryopreserved primordial germ cells in Drosophila

Miho Asaoka,Toshiyuki Takano-Shimizu,Yurina Sakamaki,Masatoshi Tomaru,Kaori Nishimura,Daisuke Tanaka,Tatsuya Fukumoto,Satoru Kobayashi

doi:10.1038/s42003-021-02692-z

Abstract

There is an urgent need to cryopreserve Drosophila stocks that have been maintained as living cultures for a long time. Long-term culture increases the risk of accidental loss and of unwanted genetic alteration. Here, we report that cryopreserved primordial germ cells (PGCs) can produce F1 progeny when transplanted into hosts. The cryopreserved donor PGCs could form germline stem cells in host gonads and contributed to continuous offspring production. Furthermore, the ability to produce offspring did not appear to vary with either differences between donor strains or cryopreservation duration. Therefore, we propose that our cryopreservation method is feasible for long-term storage of various Drosophila strains. These results underscore the potential usefulness of our cryopreservation method for backing up living stocks to avoid either accidental loss or genetic alteration.

Highlights

There is an urgent need to cryopreserve Drosophila stocks that have been maintained as living cultures for a long time
primordial germ cells (PGCs) were collected from donor embryos at the blastoderm stage using a thin glass needle and subsequently suspended in cryoprotectant agent (CPA) for cryopreservation in liquid nitrogen (LN2)
PGCs were immersed in CPAs that all contained either ethylene glycol (EG), dimethyl sulfoxide (DMSO), or glycerol (G), along with sucrose at various concentrations

Summary

Introduction

There is an urgent need to cryopreserve Drosophila stocks that have been maintained as living cultures for a long time. We found that F-PGCs transplanted into male y w hosts produced F1 progeny of both sexes (Supplementary Table 3). All females carrying GSCs differentiated from F-PGCs continued to produce donor-derived F1 progeny for at least [4,5,6] days after mating (Fig. 1e, Supplementary Fig. 2, and Supplementary Table 4).

Results

Conclusion