With the ever-expanding numbers of genetically altered (GA) animals created in this new age of CRISPR/Cas, tools for helping the management of this vast and valuable resource are essential. Cryopreservation of embryos and germplasm of GA animals has been a widely used tool for many years now, allowing for the archiving, distribution and colony management of stock. However, each year brings an array of advances, improving survival rates of embryos, success rates of in-vitro fertilisation and the ability to better share lines and refine the methods to preserve them. This article will focus on the mouse field, referencing the latest developments and assessing their efficacy and ease of implementation, with a brief note on other common genetically altered species (rat, zebrafish, Xenopus, avian species and non-human Primates).
EppigJTMotenkoHRichardsonJE, et al.The International Mouse Strain Resource (IMSR): cataloging worldwide mouse and ES cell line resources. Mamm Genome2015;
26(9–10): 448–455.
Infrafrontier Consortium.
INFRAFRONTIER–providing mutant mouse resources as research tools for the international scientific community. Nucleic Acids Res2015;
43: D1171–D1175.
6.
MobraatenLE.The Jackson Laboratory genetics stocks resource repository. Faculty Research1981. p. 219.
WhittinghamDG.Survival of mouse embryos after freezing and thawing. Nature1971;
233(5315): 125–126.
9.
WilmutI.The effect of cooling rate, warming rate, cryoprotective agent and stage of development on survival of mouse embryos during freezing and thawing. Life Sci II1972;
11(22): 1071–1079.
10.
LeiboSPMazurP.The role of cooling rates in low-temperature preservation. Cryobiology1971;
8(5): 447–452.
11.
WhittinghamDGLeiboSPMazurP.Survival of mouse embryos frozen to –196° and –269°C. Science1972;
178(4059): 411–414.
12.
NakagataN.Cryopreservation of mouse strains by ultrarapid freezing. Jikken Dobutsu1990;
39(2): 299–301.
13.
NakagataN.Cryopreservation of unfertilized mouse oocytes from inbred strains by ultrarapid freezing. Jikken Dobutsu1990;
39(2): 303–305.
14.
RallWFFahyGM.Ice-free cryopreservation of mouse embryos at –196°C by vitrification. Nature1985;
313(6003): 573–575.
15.
RallWFWoodMJKirbyC, et al.Development of mouse embryos cryopreserved by vitrification. J Reprod Fertil1987;
80(2): 499–504.
16.
KasaiM.Simple and efficient methods for vitrification of mammalian embryos. Anim Reprod Sci1996;
42(1): 67–75.
17.
KasaiM.Cryopreservation of mammalian embryos. Mol Biotechnol1997;
7(2): 173–179.
18.
KasaiMMukaidaT.Cryopreservation of animal and human embryos by vitrification. Reprod BioMed Online2004;
9(2): 164–170.
19.
PolgeCSmithAUParkesAS.Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature1949;
164(4172): 666.
20.
TadaNSatoMYamanoiJ, et al.Cryopreservation of mouse spermatozoa in the presence of raffinose and glycerol. J Reprod Fertil1990;
89(2): 511–516.
21.
YokoyamaMAkibaHKatsukiM, et al.Production of normal young following transfer of mouse embryos obtained by in vitro fertilization using cryopreserved spermatozoa. Jikken Dobutsu1990;
39(1): 125–128.
22.
SzteinJMTakeoTNakagataN.History of cryobiology, with special emphasis in evolution of mouse sperm cryopreservation. Cryobiology2018;
82: 57–63.
23.
OkuyamaMIsogaiSSagaM, et al.In-vitro fertilization (IVF) and artificial insemination (AI) by cryopreserved spermatozoa in mouse. J Fertil Implant1990;
7: 116–119.
24.
TakeshimaTNakagataNOgawaS.Cryopreservation of mouse spermatozoa. Jikken Dobutsu1991;
40(4): 493–497.
25.
SzteinJMFarleyJSYoungAF, et al.Motility of cryopreserved mouse spermatozoa affected by temperature of collection and rate of thawing. Cryobiology1997;
35(1): 46–52.
26.
OstermeierGCWilesMVFarleyJS, et al.Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation. PLoS One2008;
3(7): e2792.
27.
NakagataN.Cryopreservation of mouse spermatozoa and in vitro fertilization.Methods Mol Biol2011;
693: 57–73.
28.
TakeoTNakagataN.Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin. Biol Reprod2011;
85(5): 1066–1072.
29.
Takeo T, Nakao S, YN, Nakagawa Y, et al.Cryopreservation of mouse resources. Lab Anim Res2020;
36: 33.
30.
TakeoTNakagataN.Combination medium of cryoprotective agents containing L-glutamine and methyl-{beta}-cyclodextrin in a preincubation medium yields a high fertilization rate for cryopreserved C57BL/6J mouse sperm. Lab Anim2010;
44(2): 132–137.
31.
TakeoTHoshiiTKondoY, et al.
Methyl-beta-cyclodextrin improves fertilizing ability of C57BL/6 mouse sperm after freezing and thawing by facilitating cholesterol efflux from the cells. Biol Reprod2008;
78(3): 546–551.
32.
GuanMBoganiDMarschallS, et al.
In-vitro fertilization in mice using the MBCD-GSH protocol. Curr Protoc Mouse Biol2014;
4(2): 67–83.
33.
TakeoTNakaoSNakagawaY, et al.Cryopreservation of mouse resources. Lab Anim Res2020;
36: 33.
34.
InnaNSanmeeUSaeng-AnanU, et al.Rapid freezing versus Cryotop vitrification of mouse two-cell embryos. Clin Exp Reprod Med2018;
45(3): 110–115.
35.
ZhangJCuiJLingX, et al.
Vitrification of mouse embryos at 2-cell, 4-cell and 8-cell stages by cryotop method. J Assist Reprod Genet2009;
26(11–12): 621–628.
36.
YokotaYYokotaHYokotaM, et al.In-vitro fertilization embryo development from caffeine-treated murine sperm. Reprod Med Biol2015;
1 4(3): 117–121.
37.
Woodward B and CampbellK.Laser assisted IVF: a simplified technique to improve zygote production rates without the need for ICSI. IVF lite2014;
1(1): 12–16.
38.
HasegawaAMochidaKTomishimaT, et al.Microdroplet in-vitro fertilization can reduce the number of spermatozoa necessary for fertilizing oocytes. J Reprod Dev2014;
60(3): 187–193.
39.
NakagataNTakeoTFukumotoK, et al.
Rescue in vitro fertilization method for legacy stock of frozen mouse sperm. J Reprod Dev2014;
60(2): 168–171.
40.
TakeoTNakagataN.Superovulation using the combined administration of inhibin antiserum and equine chorionic gonadotropin increases the number of ovulated oocytes in C57BL/6 female mice. PLoS One2015;
10(5): e0128330.
41.
WangHHerathCBXiaG, et al.Superovulation, fertilization and in-vitro embryo development in mice after administration of an inhibin-neutralizing antiserum. Reproduction2001;
122(5): 809–816.
42.
TakeoTNakagataN.Cryobanking and recovery of genetically modified mice.Methods Mol Biol2020;
2066: 195–209.
43.
TakeoTMukunokiANakagataN.Ovulation of juvenile, mature, and aged female C57BL/6 mice following coadministration of inhibin antiserum and equine chorionic gonadotropin. Theriogenology2019;
135: 1–6.
44.
GotoMTakeoTTakahashiR, et al.Efficient production of immunodeficient non-obese diabetic/Shi-scid IL2rgamma(null) mice via the superovulation technique using inhibin antiserum and gonadotropin. Lab Anim2020;
55: 13–20.
45.
MeguroKKomatsuKOhdairaT, et al.
Induction of superovulation using inhibin antiserum and competence of embryo development in wild large Japanese field mice (Apodemus speciosus). Reprod Domest Anim2019;
54(12): 1637–1642.
46.
NakagataNTakeoTFukumotoK, et al.
Applications of cryopreserved unfertilized mouse oocytes for in vitro fertilization. Cryobiology2013;
67(2): 188–192.
47.
NakagawaYSakumaTSakamotoT, et al.Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. BMC Biotechnol2015;
15: 33.
48.
NakagawaYSakumaTNishimichiN, et al.
Culture time of vitrified/warmed zygotes before microinjection affects the production efficiency of CRISPR-Cas9-mediated knock-in mice. Biol Open2017;
6(5): 706–713.
49.
NakagawaYSakumaTTakeoT, et al.Electroporation-mediated genome editing in vitrified/warmed mouse zygotes created by IVF via ultra-superovulation. Exp Anim2018;
67(4): 535–543.
50.
TakeoTTsutsumiAOmaruT, et al.
Establishment of a transport system for mouse epididymal sperm at refrigerated temperatures. Cryobiology2012;
65(3): 163–168.
TakeoTFukumotoKKondoT, et al.
Investigations of motility and fertilization potential in thawed cryopreserved mouse sperm from cold-stored epididymides. Cryobiology2014;
68(1): 12–17.
53.
OkamotoMNakagataNToyodaY.Cryopreservation and transport of mouse spermatozoa at –79°C. Exp Anim2001;
50(1): 83–86.
54.
KenyonJGuanMBoganiD, et al.
Transporting mouse embryos and germplasm as frozen or unfrozen materials. Curr Protoc Mouse Biol2014;
4(2): 47–65.
55.
RaspaMFrayMPaolettiR, et al.
Long-term maintenance of frozen mouse spermatozoa at –80°C. Theriogenology2018;
107: 41–49.
56.
RaspaMFrayMPaolettiR, et al.A new, simple and efficient liquid nitrogen free method to cryopreserve mouse spermatozoa at –80°C. Theriogenology2018;
119: 52–59.
57.
MahabirEBauerBSchmidtJ.Rodent and germplasm trafficking: risks of microbial contamination in a high-tech biomedical world. ILAR J2008;
49(3): 347–355.
58.
Del ValGMRobledanoPM.In-vivo serial sampling of epididymal sperm in mice. Lab Anim2013;
47(3): 168–174.
59.
SzteinJVasudevanKRaberJ.Refinements in the cryopreservation of mouse ovaries. J Am Assoc Lab Anim Sci2010;
49(4): 420–422.
60.
DawesJLiuBMarsW, et al.Multiple ovarian transplants to rescue a transgenic line of mice. Lab Anim2010;
39(6): 191–193.
HurstJLeggettMMathersK, et al. Sharing and archiving of genetically altered mice: Opportunities for reduction and refinement. Report of the RSPCA Resource Sharing Working Group (RSWG), (2008). https://bbsrc.ukri.org/documents/genetically-altered-mice-pdf/ (n.d., accessed 13 January 2021).
66.
DickinsonMEFlennikenAMJiX, et al.
High-throughput discovery of novel developmental phenotypes. Nature2016;
537(7621): 508–514.
67.
BugeonLRosewellI.‘Mouse Locator-UK’: a networking tool for academic transgenic research in the UK.Transgenic Res2003;
12: 637.
68.
KinthPMaheshGPanwarY.Mapping of zebrafish research: a global outlook. Zebrafish2013;
10(4): 510–517.
69.
MatthewsJLMurphyJMCarmichaelC, et al.
Changes to extender, cryoprotective medium, and in-vitro fertilization improve zebrafish sperm cryopreservation. Zebrafish2018;
15(3): 279–290.
70.
Karolinska Institute of Technology. European Zebrafish Resource Center (EZRC),https://www.ezrc.kit.edu/ (n.d., accessed 13 January 2021).
71.
Institute of Hydrobiology. China Zebrafish Resource Center,http://en.zfish.cn/ (n.d., accessed 13 January 2021).
72.
LiuKPetreeCRequenaT, et al.Expanding the CRISPR toolbox in zebrafish for studying development and disease. Front Cell Dev Biol2019;
7: 13.
73.
CornetCDi DonatoVTerrienteJ.
Combining zebrafish and CRISPR/Cas9: toward a more efficient drug discovery pipeline. Front Pharmacol2018;
9: 703.
74.
CarmichaelCWesterfieldMVargaZM.Cryopreservation and in-vitro fertilization at the zebrafish international resource center.Methods Mol Biol2009;
546: 45–65.
75.
MarquesLSFossatiAANRodriguesRB, et al.
Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue. Sci Rep2019;
9(1): 15353.
76.
KhoslaKWangYHagedornM, et al.Gold nanorod induced warming of embryos from the cryogenic state enhances viability. ACS Nano2017;
11(8): 7869–7878.
77.
Caetano Da SilvaCKollarTCsenki-BakosZ, et al.an easy method for cryopreservation of zebrafish (Danio rerio) sperm. Zebrafish2019;
16(3): 321–323.
78.
MarinovicZLiQLujicJ, et al.
Preservation of zebrafish genetic resources through testis cryopreservation and spermatogonia transplantation. Sci Rep2019;
9(1): 13861.
79.
DiogoPMartinsGQuinzicoI, et al.Electric ultrafreezer (–150°C) as an alternative for zebrafish sperm cryopreservation and storage. Fish Physiol Biochem2018;
44(6): 1443–1455.
80.
GeislerRBorelNFergM, et al.Maintenance of zebrafish lines at the European Zebrafish Resource Center. Zebrafish2016;
13(Suppl 1): S19–S23.
81.
KettleboroughRNBusch-NentwichEMHarveySA, et al.
A systematic genome-wide analysis of zebrafish protein-coding gene function. Nature2013;
496(7446): 494–497.
82.
YangHTierschTR.Current status of sperm cryopreservation in biomedical research fish models: zebrafish, medaka, and Xiphophorus. Comp Biochem Physiol C Toxicol Pharmacol2009;
149(2): 224–232.
83.
SargentMGMohunTJ.Cryopreservation of sperm of Xenopus laevis and Xenopus tropicalis. Genesis2005;
41(1): 41–46.
84.
PearlEMorrowSNobleA, et al.An optimized method for cryogenic storage of Xenopus sperm to maximise the effectiveness of research using genetically altered frogs. Theriogenology2017;
92: 149–155.
85.
PearlEJGraingerRMGuilleM, et al.Development of Xenopus resource centers: the National Xenopus Resource and the European Xenopus Resource Center. Genesis. 2012;
50(3): 155–163.
Marine Biological Laboratory. National Xenopus Resource (NXR),https://www.mbl.edu/xenopus/ (2021, accessed 13 January 2021).
88.
KashiwazakiNSeitaYTakizawaA, et al.Techniques for in-vitro and in-vivo fertilization in the rat.Methods Mol Biol2010;
597: 311–322.
89.
SeitaYSugioSItoJ, et al.Generation of live rats produced by in vitro fertilization using cryopreserved spermatozoa. Biol Reprod2009;
80(3): 503–510.
90.
NakagataNMikodaNNakaoS, et al.Author Correction: establishment of sperm cryopreservation and in vitro fertilisation protocols for rats. Sci Rep2020;
10(1): 3768.
91.
HondaATachibanaRHamadaK, et al.
Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization. Sci Rep2019;
9(1): 11571.
92.
University of Missouri. Rat Resource & Research Centre (RRRC),http://www.rrrc.us/(2021, accessed 13 January 2021).
WhyteJBlesboisMcGrewM. Increased sustainability in poultry production: New tools and resources for genetic management. In: Burton E, Gatcliffe J, O’Neill HM, et al. (eds) Sustainable poultry production in Europe. Wallingford: CABI Publishing, 2016, pp. 214–234.
97.
PutkhaoKChanAWAgcaY, et al.Cryopreservation of transgenic Huntington’s disease rhesus macaque sperm: a case report. Cloning Transgenes2013;
2: 1000116.
98.
SiWZhengPTangX, et al.
Cryopreservation of rhesus macaque (Macaca mulatta) spermatozoa and their functional assessment by in-vitro fertilization. Cryobiology2000;
41(3): 232–240.
