Protocol full text hidden due to copyright restrictions
Open the protocol to access the free full text link
>
Living Beings
>
Organism
>
Specific Pathogen Free
Specific Pathogen Free
Specific Pathogn Free (SPF) is a term used to describe animals, typically laboratory animals, that are free from specific pathogens or infectious agents.
SPF animals are raised and maintained in a controlled environment to ensure they are free from specific microorganisms that could interfere with research or experimental results.
This approach is widely used in biomedical research, particularly in areas such as vaccine development, toxicology studies, and the investigation of disease mechanisms.
SPF animals provide a standardized and well-defined model system, allowing researchers to draw more reliable conclusions from their experiments and minimize the impact of confounding factors.
The use of SPF animals is a crucial aspect of ensuring the validity and reproducibility of scientific findings in the field of biomedical research.
SPF animals are raised and maintained in a controlled environment to ensure they are free from specific microorganisms that could interfere with research or experimental results.
This approach is widely used in biomedical research, particularly in areas such as vaccine development, toxicology studies, and the investigation of disease mechanisms.
SPF animals provide a standardized and well-defined model system, allowing researchers to draw more reliable conclusions from their experiments and minimize the impact of confounding factors.
The use of SPF animals is a crucial aspect of ensuring the validity and reproducibility of scientific findings in the field of biomedical research.
Most cited protocols related to «Specific Pathogen Free»
Animal Care Committees
Animals
Chimera
Mice, House
Recombinase
Rosa
Specific Pathogen Free
Strains
Tissue Donors
To generate the CD11c-Cre transgene, the 160-kb mouse genomic BAC clone RP24-361C4 (BACPAC Resources) was modified by ET recombination, as previously described (43 (link)). The clone contains the entire Itgax (CD11c) gene but lacks the 5′ end of the adjacent Itgam (CD11b) gene, preventing the overexpression of the latter. The recombination cassette containing the Cre recombinase open reading frame, followed by the bovine growth hormone (BGH) polyA signal and the FRT site-flanked prokaryotic Zeocin resistance cassette (ZeoR), replaced the coding part of the first CD11c exon, and the ZeoR cassette was subsequently removed by FLP-mediated recombination. The clone insert was released from the vector backbone using NotI digestion, gel-purified, and microinjected into fertilized oocytes. The founder line containing two copies of the transgene (as determined by quantitative Southern hybridization) was chosen for further analysis. Mice were genotyped by genomic PCR using either generic Cre primers or primers specific for the CD11c-Cre transgene (5′-ACTTGGCAGCTGTCTCCAAG-3′ and 5′-GCGAACATCTTCAGGTTCTG-3′ were specific for the CD11c promoter and Cre, respectively).
The R26-EYFP strain (21 (link)) was provided by F. Costantini (Columbia University, New York, NY). The RBP-Jfl strain (19 (link)) was provided by L. Hennighausen (National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD), with permission from T. Honjo (Kyoto University, Kyoto, Japan). The Mx1-Cre strain was previously described (44 (link)). Cre-negative RBP-Jfl/fl littermates of CKO (RBP-Jfl/fl Cre+) mice were used as controls; in preliminary experiments, wild-type CD11c-Cre+ mice were used as controls and were found indistinguishable from CD11c-Cre− animals. For inducible RBP-J deletion, adult RBP-Jfl/fl Mx1-Cre+ or control RBP-Jfl/fl mice were injected with 0.25 mg poly(I):(C) three times, with 2-d intervals, and analyzed 3 wk later. For hematopoietic reconstitution, 3 × 106 total BM cells per mouse were injected i.v. into lethally irradiated C57BL/6 mice congenic for CD45.1. The recipient mice were analyzed 4–5 wk after reconstitution. Mice were maintained in a specific pathogen-free facility and used according to the protocol approved by the Columbia University's Institutional Animal Care and Use Committee.
The R26-EYFP strain (21 (link)) was provided by F. Costantini (Columbia University, New York, NY). The RBP-Jfl strain (19 (link)) was provided by L. Hennighausen (National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD), with permission from T. Honjo (Kyoto University, Kyoto, Japan). The Mx1-Cre strain was previously described (44 (link)). Cre-negative RBP-Jfl/fl littermates of CKO (RBP-Jfl/fl Cre+) mice were used as controls; in preliminary experiments, wild-type CD11c-Cre+ mice were used as controls and were found indistinguishable from CD11c-Cre− animals. For inducible RBP-J deletion, adult RBP-Jfl/fl Mx1-Cre+ or control RBP-Jfl/fl mice were injected with 0.25 mg poly(I):(C) three times, with 2-d intervals, and analyzed 3 wk later. For hematopoietic reconstitution, 3 × 106 total BM cells per mouse were injected i.v. into lethally irradiated C57BL/6 mice congenic for CD45.1. The recipient mice were analyzed 4–5 wk after reconstitution. Mice were maintained in a specific pathogen-free facility and used according to the protocol approved by the Columbia University's Institutional Animal Care and Use Committee.
Adult
Animals
Cells
Clone Cells
Cloning Vectors
Cre recombinase
Crossbreeding
Deletion Mutation
Diabetes Mellitus
Digestion
Digestive System
Exons
Generic Drugs
Genes
Genome
growth hormone, bovine
Hematopoietic System
Institutional Animal Care and Use Committees
ITGAM protein, human
Kidney Diseases
Mice, Inbred C57BL
Mice, Laboratory
Oligonucleotide Primers
Ovum
Poly A
Poly I-C
Prokaryotic Cells
RBPJ protein, human
Recombination, Genetic
Specific Pathogen Free
Strains
Transgenes
Vertebral Column
Zeocin
Animals
Mice, House
Specific Pathogen Free
Strains
The targeting vector was constructed by insertion of a loxP sequences within introns 13 and 14 of Atg7 gene. Exon 14 was fused to a cDNA fragment encoded by exons 15, 16, and 17 (aa 1786–2097) and polyA signal sequence was added after the stop codon. Neo resistant gene cassette (mc1-neo-pA) was ligated behind the polyA signal sequence followed by the second loxP sequence, splicing acceptor site, and exon 14 with stop codon preceding the active site. We electroporated the targeting vector into mouse TT2 ES cells, selected with 200 μg/ml G418 (GIBCO BRL), and then screened for homologous recombinants by PCR and Southern blot analyses. PCR primers were as follows: 5′-TGGCTGCTACTTCTGCAATGATGT-3′, 5′-GAAGGGACTGGCTGCTATTGGGCGAAGTGC-3′, and 5′-TTAGCACAGGGAACAGCGCTCATGG-3′. Southern blot analysis was performed by digestion of genomic DNA with EcoRV and hybridization with the probe shown in Fig. 1 A. Genotyping of mice by PCR was performed using the following two primers: 5′-TGGCTGCTACTTCTGCAATGATGT-3′ and 5′-CAGGACAGAGACCATCAGCTCCAC-3′. Progeny containing the Atg7Flox allele were bred with Zp3-Cre and Mx1-Cre transgenic mice to produce Atg7−/− and Atg7F/F:Mx1 mice, respectively. With regard to Atg7F/F:Mx1 mice, Cre expression in the liver was induced by i.p. injection of pIpC (Sigma-Aldrich). 300 μl pIpC solution (1 mg/ml in water) was injected three times at 48-h intervals. Mice were housed in specific pathogen-free facilities, and the experimental protocol was approved by the Ethics Review Committee for Animal Experimentation of the Tokyo Metropolitan Institute of Medical Science.
Alleles
antibiotic G 418
Cloning Vectors
Codon, Terminator
Crossbreeding
Digestion
DNA, Complementary
Embryonic Stem Cells
Exons
Genes
Genome
Introns
Liver
Mice, Laboratory
Mice, Transgenic
Oligonucleotide Primers
Poly A
Signal Peptides
Southern Blotting
Specific Pathogen Free
The rearranged TCRα and TCRβ chains from 2D2 genomic DNA were analyzed by sequencing. The amino acid sequence of the CDR3 regions is as follows: TCRα, VYF CALRSY NFG; TCRβ, CASS LDCG ANP. The Vα3.2Jα18 and Vβ11DJβ1.1 regions of 2D2 TCR were amplified by PCR from genomic DNA with specific primers. PCR products were cloned into TCR expression cassettes (26 (link)). Linearized TCR containing plasmids were injected directly into the pronuclei of fertilized C57Bl/6 oocytes. Transgenic founders were identified by PCR using specific primers for 2D2 Vα-Jα and Vβ-Jβ regions. Transgenic founder mice were bred with C57Bl/6 mice (The Jackson Laboratory). Alternatively, 2D2 TCR transgenic mice were bred with C57Bl/6 RAG-1−/− (The Jackson Laboratory) and then intercrossed to generate 2D2 TCR transgenic RAG-1−/− mice. Routine screening to identify the transgenic mice was performed by FACS® analysis from blood using specific antibodies to Vβ11 or Vα3.2. For phenotyping, the blood was always drawn from the tail and not from the eyes of these animals. Mice were housed in a specific pathogen-free/viral antibody-free animal facility at the Harvard Institutes of Medicine. All breeding and experiments were performed in accordance with the guidelines of the committee on Animals of Harvard Medical School.
6-chloropenicillanic acid S-sulfoxide
Amino Acid Sequence
Animals
Animals, Transgenic
Antibodies
Antibodies, Viral
BLOOD
DNA Primers
Eye
Founder Mice, Transgenic
Genome
Mice, Inbred C57BL
Mice, Laboratory
Mice, Transgenic
Oligonucleotide Primers
Ovum
Plasmids
Specific Pathogen Free
Tail
T Cell Receptor beta Chain Genes
Most recents protocols related to «Specific Pathogen Free»
Example 14
In contrast to the previous experimental infection using specific pathogen-free Beagles (Crawford et al., 2005), the virus-inoculated mongrel dogs had pneumonia as evidenced by gross and histological analyses of the lungs from days 1 to 6 p.i. In addition to pneumonia, the dogs had rhinitis, tracheitis, bronchitis, and bronchiolitis similar to that described in naturally infected dogs (Crawford et al., 2005). There was epithelial necrosis and erosion of the lining of the airways and bronchial glands with neutrophil and macrophage infiltration of the submucosal tissues (
Antigens, Viral
Autopsy
Bacteria
Bronchi
Bronchioles
Bronchiolitis
Bronchitis
Canis familiaris
Epithelial Cells
Immunohistochemistry
Infection
Lung
Macrophage
Necrosis
Neutrophil
Pneumonia
Respiratory Rate
Rhinitis
Specific Pathogen Free
Superinfection
Tissues
Tracheitis
Virus
Example 1
A dose of 50 mg/kg of lipopolysaccharide (LPS) corresponds to the “lethal dose for 50 percent kill” that kills half of the population within 24 hours. Mice were subjected to intraperitoneal injection of 50 mg/kg LPS in 1×PBS for a vehicle control, and when the mice showed the signs of the moribund state, such as impaired motility, labored breathing, or inability to maintain an upright position, the mice were sacrificed by CO2 euthanasia, and the point was recorded as a humane endpoint. (The signs of the moribund state: impaired mobility, inability to maintain upright position, prolonged lack of activity and labored breathing)
All animal studies were performed according to protocols approved by Kyungpook National University (permit No. 2019-0003) and under recommendations for the proper use and care of the specific pathogen-free housing facility at Kyungpook University.
Animals
Euthanasia
Injections, Intraperitoneal
Lipopolysaccharides
Motility, Cell
Mus
Range of Motion, Articular
Sepsis
Specific Pathogen Free
Eight-week-old male C57Bl/6J mice were obtained from Wei Tong Li Hua Biotechnology Co. (Beijing, China) and housed in a specific-pathogen-free facility (22 °C, 12 h/12 h light–dark cycle) in the laboratory animal center of Tongji Medical College. All mice were allowed to adapt to the laboratory conditions for 2 weeks with free access to food and sterile water. The animal studies were approved by the Animal Ethics Committee of Tongji Medical College of Huazhong University of Science and Technology, and the works were conducted in accordance with the principles of the Animal Care and Use Committee of this institution.
Animal Ethics Committees
Animals
Animals, Laboratory
Food
Males
Mice, House
Mice, Inbred C57BL
Specific Pathogen Free
Sterility, Reproductive
Female C3H/HeJ mice 6-8-week-old were purchased from The Jackson Laboratory. All mice were housed in specific pathogen–free conditions at the Association for Assessment and Accreditation of Laboratory Animal Care Internationally Accredited University of South Carolina, School of Medicine, Animal Resource Facility and were kept under 12 light/12 dark cycles at a temperature of ∼18°C–23°C and 40%–60% humidity. Food and water were available ad libitum. All experiments performed using mice in this study were approved by the Institutional Animal Care and Use Committee, University of South Carolina under animal use protocol (AUP2363).
Animals
Animals, Laboratory
Females
Food
Humidity
Institutional Animal Care and Use Committees
Mice, House
Mice, Inbred C3H
Specific Pathogen Free
Twenty male specific-pathogen free (SPF)-grade Sprague–Dawley (SD) rats (age: 6 weeks; weight: 200 ± 20 g) were purchased from SPF (Beijing) Biotechnology Co., Ltd. Number of animal license: SYXK (Beijing) 2019–0010. This study was approved by the Ethics Committee of Beijing University of Chinese Medicine (approval number: BUCM-4-2,021,032,603-1,059).
All rats were adaptability housed for 7 days and randomly divided into four groups (n = 5 rats each) according to the method of the random number table. All rats were fed sterilized feed and deionized water in an artificial climate simulator (NHRHG6, Chongqing Hongrui Experimental Instrument Co., Ltd).
The artificial climate simulator uses a Balanced Temperature and Humidity Control (BTHC) system to control temperature (T) and relative humidity (RH). One can also design the corresponding parameters according to the actual environmental and climatic characteristics of the area and simulate the local climate. By strictly controlling environmental variable factors, the interference of other factors in experimental results can be reduced. Referring to the data1 displayed by the Spanish National Meteorological Institute (Agencia Estatal de Meteorología, AEMET), select the China-Beijing area, we established the environment temperature for the four seasons based on the information about the average monthly maximum temperature for 2020. The temperatures of the four seasons were 14.8°C in spring, 26.2°C in summer, 12.9°C in autumn, and − 2.3°C in winter. Then, the relative humidity was established with reference to the average humidity of the natural season in 2020. Accordingly, the humidity of the four seasons was 40.3% in spring, 64.3% in summer, 52.2% in autumn, and 41.2% in winter.
All rats were adaptability housed for 7 days and randomly divided into four groups (n = 5 rats each) according to the method of the random number table. All rats were fed sterilized feed and deionized water in an artificial climate simulator (NHRHG6, Chongqing Hongrui Experimental Instrument Co., Ltd).
The artificial climate simulator uses a Balanced Temperature and Humidity Control (BTHC) system to control temperature (T) and relative humidity (RH). One can also design the corresponding parameters according to the actual environmental and climatic characteristics of the area and simulate the local climate. By strictly controlling environmental variable factors, the interference of other factors in experimental results can be reduced. Referring to the data
Animals
Chinese
Climate
Ethics Committees
Hispanic or Latino
Humidity
Males
Pharmaceutical Preparations
Rattus norvegicus
Specific Pathogen Free
Top products related to «Specific Pathogen Free»
Sourced in United States, Montenegro, Germany, United Kingdom, Japan, China, Canada, Australia, France, Colombia, Netherlands, Spain
C57BL/6J is a mouse strain commonly used in biomedical research. It is a common inbred mouse strain that has been extensively characterized.
Sourced in United States, Montenegro, Canada, China, France, United Kingdom, Japan, Germany
C57BL/6 mice are a widely used inbred mouse strain commonly used in biomedical research. They are known for their black coat color and are a popular model organism due to their well-characterized genetic and physiological traits.
Sourced in China, United States, Germany, United Kingdom, Canada, Japan, France, Italy, Morocco, Spain, Netherlands, Montenegro, Belgium, Portugal, Ireland, Hungary
The C57BL/6 mouse is a widely used inbred mouse strain. It is a common laboratory mouse model utilized for a variety of research applications.
Sourced in United States, Montenegro, United Kingdom, Germany, Australia, China, Canada
C57BL/6 is a widely used inbred mouse strain. It is a robust, readily available laboratory mouse model.
Sourced in United States, Montenegro, Japan, Canada, United Kingdom, Germany, Macao, Switzerland, China
C57BL/6J mice are a widely used inbred mouse strain. They are a commonly used model organism in biomedical research.
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
Sourced in China, United States, Germany, Japan, Canada, United Kingdom, France, Italy, Spain
BALB/c mice are an inbred strain of laboratory mice commonly used in scientific research. They are a widely utilized model organism for various experiments and studies. The BALB/c strain is known for its susceptibility to certain diseases and its ability to produce high levels of antibodies, making it a valuable tool for immunological research.
Sourced in China, Japan, Germany, France, United Kingdom, United States, Italy, Canada, Montenegro, Belgium, Morocco, Netherlands, Spain
The C57BL/6J mouse is a widely used laboratory mouse strain. It is an inbred strain that has a black coat color. The C57BL/6J mouse is commonly used as a control strain in various research applications.
Sourced in China, Japan, United States, Germany, United Kingdom, France, Italy
BALB/c nude mice are an inbred strain of mice that lack a functional immune system due to a genetic mutation. They are athymic, meaning they lack a thymus gland, which is essential for the development of T cells. This results in a severely compromised adaptive immune response. BALB/c nude mice are commonly used in biomedical research for the study of human diseases, the evaluation of new therapies, and the development of xenograft models.
Sourced in United States, China, United Kingdom, Germany, France, Australia, Canada, Japan, Italy, Switzerland, Belgium, Austria, Spain, Israel, New Zealand, Ireland, Denmark, India, Poland, Sweden, Argentina, Netherlands, Brazil, Macao, Singapore, Sao Tome and Principe, Cameroon, Hong Kong, Portugal, Morocco, Hungary, Finland, Puerto Rico, Holy See (Vatican City State), Gabon, Bulgaria, Norway, Jamaica
DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
More about "Specific Pathogen Free"
Specific Pathogen Free (SPF) is a crucial concept in biomedical research, ensuring the validity and reproducibility of scientific findings.
SPF animals, typically laboratory animals like mice and rats, are raised and maintained in a controlled environment to be free from specific infectious agents or pathogens that could interfere with research.
This approach is widely used in areas such as vaccine development, toxicology studies, and disease mechanism investigations.
The use of SPF animals provides a standardized and well-defined model system, allowing researchers to draw more reliable conclusions and minimize the impact of confounding factors.
Common SPF animal models include the C57BL/6J and BALB/c mouse strains, which are extensively used in biomedical research.
These mice are often supplemented with Fetal Bovine Serum (FBS) and cultured in Dulbecco's Modified Eagle Medium (DMEM) to ensure optimal growth and health.
Streamlining SPF research protocols is crucial, and platforms like PubCompare.ai can help researchers effortlessly locate and compare the best protocols from literature, pre-prints, and patents.
By leveraging AI-driven comparisons, researchers can identify the most suitable protocols and products for their SPF research needs, optimizing their workflow and enhancing the overall quality of their studies.
Whether you're working with SPF mice, rats, or other laboratory animals, understanding the importance of this approach and utilizing the right tools and resources can lead to more robust and reliable scientific findings in the field of biomedical research.
With the power of SPF and the support of innovative platforms like PubCompare.ai, researchers can unlock new insights and push the boundaries of scientific discovery.
SPF animals, typically laboratory animals like mice and rats, are raised and maintained in a controlled environment to be free from specific infectious agents or pathogens that could interfere with research.
This approach is widely used in areas such as vaccine development, toxicology studies, and disease mechanism investigations.
The use of SPF animals provides a standardized and well-defined model system, allowing researchers to draw more reliable conclusions and minimize the impact of confounding factors.
Common SPF animal models include the C57BL/6J and BALB/c mouse strains, which are extensively used in biomedical research.
These mice are often supplemented with Fetal Bovine Serum (FBS) and cultured in Dulbecco's Modified Eagle Medium (DMEM) to ensure optimal growth and health.
Streamlining SPF research protocols is crucial, and platforms like PubCompare.ai can help researchers effortlessly locate and compare the best protocols from literature, pre-prints, and patents.
By leveraging AI-driven comparisons, researchers can identify the most suitable protocols and products for their SPF research needs, optimizing their workflow and enhancing the overall quality of their studies.
Whether you're working with SPF mice, rats, or other laboratory animals, understanding the importance of this approach and utilizing the right tools and resources can lead to more robust and reliable scientific findings in the field of biomedical research.
With the power of SPF and the support of innovative platforms like PubCompare.ai, researchers can unlock new insights and push the boundaries of scientific discovery.