B. infantis ATCC 15697 (American Type Culture Collection) was grown from a frozen stock in deMan, Rogosa, Sharpe broth (Difco) supplemented with cysteine (0.05%) and incubated anaerobically as previously described (20 (link)). Midexponential phase B. infantis was harvested by centrifugation, resuspended in sterile PBS, and cryogenically preserved in a 1:1 cell to glycerol (25%) suspension. Each batch of B. infantis inoculum was validated for viability on Reinforced Clostridial Agar. On average, each stock contained 5 × 108 CFUs/mL. Before administration, B. infantis was washed in PBS, and the bacterial pellet was resuspended in PBS for administration. Piglets in the BI and COMB groups were orally gavaged with 2 mL washed B. infantis 3 times daily for a total dosage of ∼3 × 109 CFUs/d. This dose was selected on the basis of typical probiotic dosing regimens (21 (link)–23 (link)).
Atcc 15697
Manufactured by American Type Culture Collection
Sourced in United Kingdom
ATCC® 15697™ is a cryopreserved cell line derived from the human cell line HeLa. It is a widely used and well-characterized cell line for various research applications.
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Lab products found in correlation
3 protocols using atcc 15697
1
Cultivation and Dosing of B. infantis ATCC 15697
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B. infantis ATCC 15697 (American Type Culture Collection) was grown from a frozen stock in deMan, Rogosa, Sharpe broth (Difco) supplemented with cysteine (0.05%) and incubated anaerobically as previously described (20 (link)). Midexponential phase B. infantis was harvested by centrifugation, resuspended in sterile PBS, and cryogenically preserved in a 1:1 cell to glycerol (25%) suspension. Each batch of B. infantis inoculum was validated for viability on Reinforced Clostridial Agar. On average, each stock contained 5 × 108 CFUs/mL. Before administration, B. infantis was washed in PBS, and the bacterial pellet was resuspended in PBS for administration. Piglets in the BI and COMB groups were orally gavaged with 2 mL washed B. infantis 3 times daily for a total dosage of ∼3 × 109 CFUs/d. This dose was selected on the basis of typical probiotic dosing regimens (21 (link)–23 (link)).
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B. infantis ATCC 15697 (American Type Culture Collection) was grown from a frozen stock in deMan, Rogosa, Sharpe broth (Difco) supplemented with cysteine (0.05%) and incubated anaerobically as previously described (20 (link)). Midexponential phase B. infantis was harvested by centrifugation, resuspended in sterile PBS, and cryogenically preserved in a 1:1 cell to glycerol (25%) suspension. Each batch of B. infantis inoculum was validated for viability on Reinforced Clostridial Agar. On average, each stock contained 5 × 108 CFUs/mL. Before administration, B. infantis was washed in PBS, and the bacterial pellet was resuspended in PBS for administration. Piglets in the BI and COMB groups were orally gavaged with 2 mL washed B. infantis 3 times daily for a total dosage of ∼3 × 109 CFUs/d. This dose was selected on the basis of typical probiotic dosing regimens (21 (link)–23 (link)).
2
Protein Determination and Bacterial Strains
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The IGEP was kindly provided by Upfront Chromatography (Copenhagen, Denmark). The protein content of the powder was determined by the Bradford assay [20 (link)]. B. longum subsp. infantis ATCC® 15697™ (B. infantis) was obtained from the American Type Culture Collection (ATCC, Middlesex, UK). C. jejuni 81–176 was kindly provided by Dr Marguerite Clyne’s, University College Dublin.
3
Bifidobacterium longum subsp. infantis Culture
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Bacterial culture conditions were maintained as previously described [9 (link)]. Bifidobacterium longum subsp. infantis ATCC® 15697™ (B. infantis) was obtained from the American Type Culture Collection (ATCC, Middlesex, UK). and the strain was stored in deMan Rogosa Sharpe (MRS) (Difco, Sparks, MD, USA) broth containing 50% glycerol at −80 °C. The strain was cultured twice in MRS media supplemented with L-cysteine (0.05% w/v) prior to use and was routinely grown overnight at 37 °C under anaerobic conditions generated using the Anaerocult A system (Merck, Dannstadt, Germany). For assays, B. infantis was incubated alone or in combination with each powder for 1 h and the bacteria were serially diluted and enumerated by spot plating on MRS agar to determine bacterial numbers. To determine the mid-exponential growth phase, the culture was monitored by measuring the absorbance at 600 nm (OD600 nm) at intervals during growth.
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