F. nucleatum (ATCC 25586), P. anaerobius (ATCC 27337), and P. micra (ATCC 33270) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). All bacteria were cultured at 37°C in Anaerobe basal broth (CM0957, OXOID, Thermo Fisher Scientific, West Palm Beach, FL, USA) in an anaerobic jar (Hardy Diagnostics, Santa Maria, CA, USA). Anaerobic conditions were maintained using AnaeroGen packets (Thermo Fisher Scientific).
F nucleatum
Manufactured by American Type Culture Collection
Sourced in United States
F. nucleatum is a bacterial culture available from the American Type Culture Collection. It is an anaerobic, non-spore-forming, gram-negative bacterium. The core function of this culture is to serve as a research tool for scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
P gingivalis, by American Type Culture Collection (4 mentions)
P micra, by American Type Culture Collection (2 mentions)
S mitis, by American Type Culture Collection (2 mentions)
Hemin, by Merck Group (2 mentions)
S mutans, by American Type Culture Collection (2 mentions)
Anaerobic jar, by Hardy Diagnostics (1 mentions)
Anaerobe basal broth, by Thermo Fisher Scientific (1 mentions)
S oralis, by American Type Culture Collection (1 mentions)
F nucleatum, by Merck Group (1 mentions)
Bhi broth, by Acumedia (1 mentions)
Bhi broth, by Merck Group (1 mentions)
Anaerobic chamber, by Don Whitley Scientific (1 mentions)
S gordonii, by American Type Culture Collection (1 mentions)
T denticola, by American Type Culture Collection (1 mentions)
S salivarius, by American Type Culture Collection (1 mentions)
L rhamnosus, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
11 protocols using f nucleatum
1
Culturing Anaerobic Oral Bacteria
2
Culturing Oral Bacterial Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
S. mitis (ATCC 49456), S. oralis (ATCC 35037),
and F. nucleatum (ATCC 25586) strains were purchased from the
American Type Culture Collection. S. mitis and S.
oralis were cultured in brain heart infusion (BHI) broth (Acumedia)
in a humidified incubator with 5% CO2 at 37°C. F.
nucleatum was cultured in BHI broth supplemented with 0.5% yeast
extract, 5 µg/mL hemin (Sigma-Aldrich), and 1 µg/mL vitamin K, and incubated at
37°C in an anaerobic chamber (Don Whitley Scientific).
and F. nucleatum (ATCC 25586) strains were purchased from the
American Type Culture Collection. S. mitis and S.
oralis were cultured in brain heart infusion (BHI) broth (Acumedia)
in a humidified incubator with 5% CO2 at 37°C. F.
nucleatum was cultured in BHI broth supplemented with 0.5% yeast
extract, 5 µg/mL hemin (Sigma-Aldrich), and 1 µg/mL vitamin K, and incubated at
37°C in an anaerobic chamber (Don Whitley Scientific).
3
Anaerobic Bacterial Culture and LPS Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anaerobic bacteria T. denticola (ATCC 35405), P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) were purchased from ATCC. T. denticola, P. gingivalis and F. nucleatum were grown as described previously at 37°C under oxygen deprived anaerobic conditions in New Oral Spirochete Medium or in Brain-heart Infusion (BHI) broth supplemented with hemin (5 μg ml-1) and vitamin K (1 μg ml-1) [143 , 144 (link)]. Purity of spirochete cultures was confirmed by dark field microscopy prior to use in experiments. Oral commensal bacteria S. gordonii (ATCC 10558), S. salivarius (ATCC 13419) and V. parvula (ATCC 10790) were grown overnight at 37°C under anaerobic conditions in BHI broth then harvested for subsequent assays. Gram staining followed by microscopic evaluation and colony morphology on Mitis Salivarius plates were used to confirm purity of cultures. T.denticola lipoologosaccharide (Td-LOS) was a gift from Daniel Grenier, Universite Laval, Quebec, Canada, P.gingivalis lipopolysaccharide (Pg-LPS) and F.nucleatum lipopolysaccharide (Fn-LPS) were provided by Richard P. Darveau, University of Washington, Seattle and V. parvula lipopolysaccharide (Vp-LPS) was provided by Christopher Fenno, University of Michigan, Ann Arbor, MI.
4
Caco-2 Cell Responses to F. nucleatum and L. rhamnosus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human colonic adenoma cell line, Caco-2, was purchased from the American Type Culture Collection (ATCC) and cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.), with 10% fetal bovine serum (FBS, Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml streptomycin/penicillin (Gibco; Thermo Fisher Scientific, Inc.) and 1% HEPES (Gibco; Thermo Fisher Scientific, Inc.) at 37°C under 5% CO2. F. nucleatum (ATCC 25586) and L. rhamnosus (ATCC 11982) were purchased from ATCC and cultured by the Wuhan Research Institute of First Light Industry (Wuhan, China). The methods of bacterial pellets and conditioned medium were as previously described (19 (link)). Caco-2 cells were treated with the supernatant of F. nucleatum and/or L. rhamnosus to examine the effects of F. nucleatum and L. rhamnosus (20 (link),21 (link)). 3-MA (5 mM) and CQ (10 µM) treatment for 2 h was used to inhibit the autophagic flux.
5
Oral Microbiome Strain Collection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following strains commonly present in the oral microbiome (Aas et al., 2005 (link); Maddi and Scannapieco, 2013 (link); Loozen et al., 2014 (link)) were obtained from the American Type Culture Collection (ATCC): A. actinomycetemcomitans (ATCC 43718), F. nucleatum (ATCC 10953), P. gingivalis (ATCC 33277), P. intermedia (ATCC 25611), S. mutans (ATCC 25175), S. sobrinus (ATCC 33478), T. forsythia (ATCC 43037), Actinomyces naeslundii (ATCC 51655), Capnocytophaga sputigena (ATCC 33612), Streptococcus gordonii (ATCC 49818), Actinomyces viscosus (ATCC 15987), and S. mitis (ATCC 49456). Veillonella parvula was obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures (DSM 2007); S. sanguinis was acquired from the BCCM/LMG Bacteria Collection (LMG 14657). S. salivarius strain TOVE-R (Tanzer et al., 1985 (link)) was utilized. Bacterial strains were classified as either commensal (indigenous), potentially health-associated or disease-associated, as indicated in Table 1 .
6
Standardized Microbial Culture Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reference strains from American Type Culture Collection (ATCC) were used such as C. albicans (ATCC 18804), E. faecalis (ATCC 4083), P. aeruginosa (ATCC 15442), S. aureus (ATCC 6538), S. mutans (ATCC 35688), F. nucleatum (ATCC 25586), P. endodontalis (ATCC 35406), P. gingivalis (ATCC 33277), and P. micra (ATCC 23195). The microorganisms were frozen at -80 °C, and activated in a specific culture medium according to the metabolic needs, being Yeast Nitrogen Base broth (YNB -Sigma-Aldrich, St. Louis, USA), for C. albicans, and Brain Heart Infusion broth (BHI -Himedia), for aerobic bacteria. They were incubated at 37 °C for 24 h, with 5% CO2 for S. mutans. For anaerobic bacteria, Brucella broth (Acumedia, Michigan, USA) supplemented with 5% sterile defibrinated sheep blood (Newprov, Pinhais-PR, Brazil), 1% hemin (Sigma-Aldrich), and 1% menadione (Sigma-Aldrich) was used. These bacteria were incubated in anaerobiosis with nitrogen 80.01%, carbon dioxide 10.02%, and hydrogen 9.97%; (Whitley DG250 Workstation, West Yorkshire, UK) at 37 ºC for 48 h.
7
Cultivation and Maintenance of Oral Bacteria
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lactobacillus reuteri strains KCTC 3594, KCTC 3678, KCTC 3679, KCTC 3680, KCTC 3682, and KCTC 3683 were used as reference strains. All samples were purchased from the Korean Collection for Type Cultures (KCTC, Daejeon, Korea). Reference strains and newly isolated L. reuteri strains (AN417, AN306, AN403, AN413, AN507, AN509, AN510, AN511, AN513, AN519, AN523, AN705, AN711, and RI-7) were grown on MRS agar plates. P. gingivalis strain BAA-308 and F. nucleatum KCTC 15573 were purchased from the American Type Culture Collection (Manassas, VA, USA) and KCTC, respectively. They were grown in trypticase soy broth (TSB; BD, Germany) comprised of (per L): 30 g trypticase soy broth, 5 mg hemin, 5 g yeast extract, 1 mg vitamin K1, and 15 g agar, or blood TSB agar (TSB medium plus 15 g/L agar and supplemented with 3% sheep blood). For all experiments, P. gingivalis and F. nucleatum were also cultured in TSB broth for at least 12 h prior to inoculation. Bacteria were grown and maintained at 37 °C in an anaerobic chamber in an atmosphere of CO2:H2:N2 (5:10:85). S. mutans (KCTC 3065) strains were grown in brain heart infusion (BHI) medium at 37 °C under aerobic conditions.
8
Cultivation and Enumeration of Fusobacterium nucleatum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial strain F. nucleatum (ATCC 25586) was obtained from the American Type Culture Collection (ATCC). The strain was cultured on Brucella agar plates with 5% sheep blood and incubated in a Coy chamber (Coy Laboratory Product, USA) for 3 days at 37°C under anaerobic conditions (atmosphere: 20% CO2, 5% H2, and 75% N2). A single colony was inoculated into a Gifu anaerobic medium (GAM) broth (KisanBio, Korea) and incubated at 37°C for 18 hr. Bacterial cells were centrifuged at 2,000 х g for 5 min at 4°C and washed twice with PBS. For calculating colony-forming units per millimeter (CFU/mL) of a fluid, the pellet was serially diluted 10-fold in fresh GAM broth and spread onto the Brucella agar plate. The mean colony counts were 1.0 x 109 CFU/mL.
9
Cultivation of Gut Microbiome Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
F.nucleatum, A.muciniphila and B.adolescentis were purchased from American Type Culture Collection (ATCC, F.nucleatum strain 25586 and 10953, A.muciniphila BAA-835 and B.adolescentis strain 15703). F.nucleatum was grown in Columbia blood agar (Comagal, China) in an anaerobic glove box at 37°C. A.muciniphila in Brain Heart Infusion (BD Difco, USA) supplemented with 0.05% mucin Type II (Sigma-Aldrich, USA), B.adolescentis in Reinforced Clostridium Medium (BD Difco, USA) were cultured under an atmosphere of 10% H2, 10% CO2, and 80% N2 in an AW500SG anaerobic workstation (ELECTROTEK) at 37°C. The E.coli strain DH5α (Takara, Japan) was cultured in Luria-Bertani medium overnight at 37°C.
10
Culturing Oral Pathogens and E. coli
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
P. gingivalis used in this study 53978 (W50), BAA-308 (W83), ATCC33277, and F. nucleatum, were obtained from American Type Culture Collection (ATCC). P. gingivalis W50-NIDCR was a generous gift from Dr. P. Kolenbrander in the National Institute of Dental and Craniofacial Research (NIDCR). All oral bacteria were grown anaerobically (GasPak EZ; BD, Sparks, MD) in BactoTM Brain Heart Infusion (BD, Sparks, MD or Anaerobe Systems Morgan Hill, CA), supplemented with 5 mg/L hemin and 1 mg/L vitamin K. E. coli were grown in Luria Bertani Broth (K.D Medical Columbia, MD). Bacteria were harvested in the exponential stage of their growth and washed 2x with PBS before being used at an O.D. of 600 nm.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!