Cronobacter sakazakii strains (strain no. ATCC29544, ATCC12868, ATCC29004, and ATCCBAA-894) and human brain microvascular endothelial cells (HBMEC) were purchased from the American Type Culture Collection (ATCC). Bacterial strains were grown at 37°C in brain heart infusion (BHI) broth (Landbridge, Beijing, China) overnight. HBMEC were grown in DMEM (Dulbecco’s modified eagle medium, Gibco, Grand Island, NY, United States) with 10% FBS (Biological Industries, Kibbutz Beit Haemek, Israel) at 37°C and 5% CO2 for the indicated period of time in different experiments.
Atcc 29544
Manufactured by American Type Culture Collection
Sourced in United States
ATCC 29544 is a laboratory equipment product offered by American Type Culture Collection. It is a microbiological growth medium used for the cultivation and maintenance of bacterial cultures. The product provides a standardized and controlled environment for the propagation of various bacterial species.
Automatically generated - may contain errors
Lab products found in correlation
Atcc 29004, by American Type Culture Collection (4 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Hbmec, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Atcc12868, by American Type Culture Collection (1 mentions)
Atccbaa 894, by American Type Culture Collection (1 mentions)
Hbmec, by American Type Culture Collection (1 mentions)
6 protocols using atcc 29544
1
Cronobacter sakazakii Infection of HBMEC
2
Coenzyme Q10 Inhibits Cronobacter Biofilm
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CoQ 0 (HPLC ≥99%, CAS 605-94-7) was obtained from J&K Scienti c Co., Ltd (Beijing, China) and dissolved in dimethyl sulfoxide (DMSO) for use in all experiments. The nal concentration of DMSO in all sample solutions was 0.1% (v/v), which has no apparent effect on the growth of C. sakazakii. All other chemicals were of analytical grade and were unaltered.
Bacterial strains and culture conditions C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong bio lm producer, was used in the bio lm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
Bacterial strains and culture conditions C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong bio lm producer, was used in the bio lm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
3
Isolation and Activation of Cronobacter sakazakii
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Three strains of C. sakazakii (ATCC 29544, 14-15, and 18-8) were used for this study. ATCC 29544 was procured from the American Type Culture Collection (ATCC, Manassas, VA, United States). Strains 14-15 and 18-8 were isolated from PIF and baby formula, respectively (Li et al., 2016 (link)). All C. sakazakii strains were stored at -80°C in Luria-Bertani (LB) broth with 30% (vol/vol) glycerin. To activate the frozen cultures, a loopful of each strain was streaked with a flamed loop onto Tryptic Soy Agar (TSA, Land Bridge, Beijing, China) and incubated at 37°C for 24 h, followed by incubation at 37°C in sterile Tryptic Soy Broth (TSB; Land Bridge) for 18 h.
4
Cultivation of C. sakazakii CECT 858
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The bacterial strain used in this study was C. sakazakii CECT 858, supplied by the Spanish Type Culture Collection (CECT, Valencia, Spain), which corresponds with the strain ATCC 29544 of the American Type Culture Collection and is of clinical origin from a child throat. It is recommended as a reference strain by international standards ISO 22964:2017 and ISO 11133:2014/Amd 2:2020. For the reference stock, the bacteria were fixed to porous rings and stored in cryovials at − 80 °C. To cultivate C. sakazakii, we followed the procedure described in our previous study (Abad et al. 2022 (link)).
5
Cultivation and Standardization of C. sakazakii
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C. sakazakii strains ATCC 29004 and ATCC 29544 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Strain ATCC 29004, which is a relatively strong biofilm producer, was used in the biofilm assay. Prior to each assay, bacteria were inoculated onto tryptic soy agar (TSA) medium and incubated at 37°C for 12 h. To obtain fresh overnight cultures, a single colony was inoculated into 30 mL of tryptic soy broth (TSB) medium and incubated with shaking at 130 rpm for 12 h at 37°C. Following incubation, cultures were centrifuged (4°C, 8000 × g, 5 min), washed three times with sterile phosphate-buffered saline (PBS), and diluted in TSB medium to an optical density at 600 nm (OD 600 ) of 0.5 (approximately 4 × 10 8 colony-forming units (CFU)/mL).
6
Characterization of Cronobacter sakazakii Strains
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C. sakazakii strains ATCC 29544 and ATCC 29004 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Three other C. sakazakii strains (CS 1, CS 2, CS 3) were taken from our laboratory strain collection, which were originally isolated from infant formula and infant rice cereal collected from supermarkets in China. All isolates were used in minimum inhibitory concentration assay and only C. sakazakii ATCC 29544 was used for antimicrobial mechanism analysis. All strains were stored in tryptone soya broth (TSB) with 20% glycerol (v/v) at -80°C. Before each experiment, stock cultures were streaked on tryptone soya agar (TSA) and grew at 37°C for 18 h. Then a loopful of each strain was inoculated into 30 mL TSB and incubated for 18 h at 37°C.
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