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Mouse Sperm Freezing
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For laboratory animals, in particular, animal models, sperm freezing is a very secure and cost-efficient conservation approach; in Haimen Animal Center, the fertilization rate of thawed frozen sperms is able to be kept above 30% or at an even higher level, and for C57BL/6 mice in particular, the thawing rate of frozen sperms can be above 50%.
Mouse Sperm Freezing Flow Chart:
Specification for Mouse Sperm Freezing
Acceptance Criteria for Parental Mice
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- Parental mice to be supplied by the client: 3–5 fertile male mice (10–20 weeks of age).
- Documents to be filled out by the client:
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- Genotype identification protocol for the mice to be frozen (Biocytogen BD will send the protocol to the client)
Delivery Specification for Frozen Sperms
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- Quantity of delivered straws: 20 straws/study.
- Quantity of delivered mice: It is warranted that not fewer than 10 viable animals can be obtained from each straw when thawed in Biocytogen depending on the customer’s target.
- Genotype of delivered mice: Homozygous/heterozygous depending on the customer’s target.
- Microbial level of delivered mice: The mice are tested negative for all 45 tests (see Table 2).
- Accompanying documents submitted at the time of mouse delivery:
- Mouse genotype identification report;
- Microbial testing report for mice (An analysis report for corresponding tests will be provided upon request of the client, and testing cost will be charged separately depending on test specifics).
Table 2. Pathogen Exclusion List in Biocytogen Animal Facility
No. Test Item 1 Salmonella spp. 2 Corynebacterium kutscheri 3 Pasteurella pneumotropica 4 Klebsiella pneumoniae 5 Staphylococcus aureus 6 Pseudomonas aeruginosa 7 Ectoparasites 8 Ectromelia Virus 9 Mouse Hepatitis Virus (MHV) 10 Sendai Virus 11 Pneumonia Virus of Mice (PVM) 12 Reovirus 3 (Reo 3) 13 Minute Virus of Mice 14 Clostridium piliforme 15 Lymphocytic Choriomeningitis Virus 16 Hantaan Virus 17 Mouse Encephalomyelitis Virus(GDVII)/Rat Theilovirus (RTV) 18 Mouse Adenovirus I and II (FL and K88) 19 Polyoma Virus 20 Streptococcus pneumoniae 21 Encephalitozoon cuniculi 22 Beta hemolytic Streptococcus (non-Group D) 23 Mycoplasma spp 24 Mycoplasma Pulmonis 25 Enteric Helminths 26 Endoparasites 27 Ciliates 28 Toxoplasma 29 Bordetella bronchiseptica 30 Pasteurella multocida 31 Klebsiella oxytoca 32 Citrobacter rodentium 33 Proteus spp. 34 Mouse Parvovirus (MPV) 35 Murine Norovirus (MNV) 36 Mouse Rotavirus (EDIM) 37 K Virus 38 Mouse Cytomegalovirus (MCMV) 39 Cilia Associated Respiratory Bacillus 40 Thymic Virus 41 Lactate Dehydrogenase Elevating Virus 42 Helicobacter spp. 43 Streptococcus B-hemolytic (Groups B) 44 Streptococcus B-hemolytic (Groups G) 45 NS-1 -
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Thawing of Frozen Mouse Sperms
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Frozen sperms provided by the client can be thawed in Biocytogen’s Haimen Animal Center, too, followed by embryo transfer by Biocytogen’s IVF team in the high-standard animal room to result in mice of SPF or even higher grade.
Thawing of Frozen Mouse Sperms Service Flowchart:
Specification for Frozen Mouse Sperm Thawing Services
Acceptance Criteria for Frozen Sperms
- Straws to be provided by the client: >5 straws/study.
- Documents to be filled out by the client:
- Genotype identification protocol for frozen mice (Biocytogen BD will send the protocol to the client)
Delivery Specification for Thawed Frozen Sperms
- Quantity of delivered mice: It is warranted that not fewer than 10 viable animals can be obtained from each straw, depending on the customer’s target.
- Genotype of delivered mice: Consistent with the genotype required by the client.
- Microbial level of delivered mice: The mice are tested negative for all 45 tests (see Table 2).
- Accompanying documents submitted at the time of mouse delivery:
- Mouse genotype identification report;
- Microbial testing report for mice (An analysis report for corresponding tests will be provided upon request of the client, and testing cost will be charged separately depending on test specifics).
Table 2. Pathogen Exclusion List in Biocytogen Animal Facility
No. Test Item 1 Salmonella spp. 2 Corynebacterium kutscheri 3 Pasteurella pneumotropica 4 Klebsiella pneumoniae 5 Staphylococcus aureus 6 Pseudomonas aeruginosa 7 Ectoparasites 8 Ectromelia Virus 9 Mouse Hepatitis Virus (MHV) 10 Sendai Virus 11 Pneumonia Virus of Mice (PVM) 12 Reovirus 3 (Reo 3) 13 Minute Virus of Mice 14 Clostridium piliforme 15 Lymphocytic Choriomeningitis Virus 16 Hantaan Virus 17 Mouse Encephalomyelitis Virus(GDVII)/Rat Theilovirus (RTV) 18 Mouse Adenovirus I and II (FL and K88) 19 Polyoma Virus 20 Streptococcus pneumoniae 21 Encephalitozoon cuniculi 22 Beta hemolytic Streptococcus (non-Group D) 23 Mycoplasma spp 24 Mycoplasma Pulmonis 25 Enteric Helminths 26 Endoparasites 27 Ciliates 28 Toxoplasma 29 Bordetella bronchiseptica 30 Pasteurella multocida 31 Klebsiella oxytoca 32 Citrobacter rodentium 33 Proteus spp. 34 Mouse Parvovirus (MPV) 35 Murine Norovirus (MNV) 36 Mouse Rotavirus (EDIM) 37 K Virus 38 Mouse Cytomegalovirus (MCMV) 39 Cilia Associated Respiratory Bacillus 40 Thymic Virus 41 Lactate Dehydrogenase Elevating Virus 42 Helicobacter spp. 43 Streptococcus B-hemolytic (Groups B) 44 Streptococcus B-hemolytic (Groups G) 45 NS-1 -
Mouse Embryo Freezing
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Embryos of genetically edited animal models can be frozen rapidly by multiple ovulation, IVF or natural co-caging and embryo flushing; thereby, provided that the parental mice are not sterilized, animal feeding cost is reduced, and the probability of genetic drift in prolonged conservation of viable organisms is lower, too. In contrast to sperm freezing, homozygous embryos are normally frozen in embryo breezing, hence the mice of the target genotype can be obtained rapidly after embryo thawing.
Mouse Embryo Freezing Service Flowchart
Specification for Mouse Embryo Freezing Services
Acceptance Criteria for Parental Mice
- Quantity of animals to be provided by the client: Heterozygous species conservation: 3-5 fertile male mice; homozygous species conservation: 3-5 fertile homozygous male mice, and >=25 homozygous female mice.
- Ages of animals provided by the client: male mice: 10–20 weeks, female mice: 4–12 weeks.
- Documents to be filled out by the client
- Genotype identification protocol for mice to be frozen (Biocytogen BD will send the protocol to the client);
Delivery Specification for Frozen Mouse Embryos
- Quantity of delivered mice: It is warranted that not fewer than 10 viable animals can be obtained from each straw, depending on the customer’s target.
- Genotype of delivered mice: Consistent with the genotype required by the client.
- Microbial level of delivered mice: The mice are tested negative for all 45 tests (see Table 2).
- Accompanying documents submitted at the time of mouse delivery:
- Mouse genotype identification report;
- Microbial testing report for mice (An analysis report for corresponding tests will be provided upon request of the client, and testing cost will be charged separately depending on test specifics).
Table 2. Pathogen Exclusion List in Biocytogen Animal Facility
No. Test Item 1 Salmonella spp. 2 Corynebacterium kutscheri 3 Pasteurella pneumotropica 4 Klebsiella pneumoniae 5 Staphylococcus aureus 6 Pseudomonas aeruginosa 7 Ectoparasites 8 Ectromelia Virus 9 Mouse Hepatitis Virus (MHV) 10 Sendai Virus 11 Pneumonia Virus of Mice (PVM) 12 Reovirus 3 (Reo 3) 13 Minute Virus of Mice 14 Clostridium piliforme 15 Lymphocytic Choriomeningitis Virus 16 Hantaan Virus 17 Mouse Encephalomyelitis Virus(GDVII)/Rat Theilovirus (RTV) 18 Mouse Adenovirus I and II (FL and K88) 19 Polyoma Virus 20 Streptococcus pneumoniae 21 Encephalitozoon cuniculi 22 Beta hemolytic Streptococcus (non-Group D) 23 Mycoplasma spp 24 Mycoplasma Pulmonis 25 Enteric Helminths 26 Endoparasites 27 Ciliates 28 Toxoplasma 29 Bordetella bronchiseptica 30 Pasteurella multocida 31 Klebsiella oxytoca 32 Citrobacter rodentium 33 Proteus spp. 34 Mouse Parvovirus (MPV) 35 Murine Norovirus (MNV) 36 Mouse Rotavirus (EDIM) 37 K Virus 38 Mouse Cytomegalovirus (MCMV) 39 Cilia Associated Respiratory Bacillus 40 Thymic Virus 41 Lactate Dehydrogenase Elevating Virus 42 Helicobacter spp. 43 Streptococcus B-hemolytic (Groups B) 44 Streptococcus B-hemolytic (Groups G) 45 NS-1
