MC3T3-E1 preosteoblasts[30 (link)] (RIKEN Cell Bank, Tsukuba, Japan) were cultured in regular culture media consisting of alpha-modified minimum essential medium (alpha-MEM; Nacalai Tesque) supplemented with 10% heat-inactivated fetal bovine serum (GIBCO, Grand Island, NY, USA) and 1% Antibiotic‐Antimycotic Mixed Stock Solution (Nacalai Tesque) in a humidified atmosphere of 5% CO2 at 37°C. The cells were trypsinized and seeded onto sample scaffolds at 4 x 104cell /scaffold before each experiment. In all experiments, cells (20 μL of each cell suspension at a suitable cell concentration) were added to each scaffold using a micropipette and incubated in a 5% CO2 saturated humidity incubator for an hour at 37°C until the cells adhered to the scaffold. Thereafter, each scaffold was transferred to a plate (size suitable for each experiment) and cultured in an appropriate amount of alpha-MEM with medium replacement twice a week.
Antibiotic antimycotic mixed stock solution
Manufactured by Nacalai Tesque
Sourced in Japan, United States, France
Antibiotic-antimycotic mixed stock solution is a sterile liquid containing a combination of antibiotics and antifungal agents. It is commonly used in cell culture applications to prevent bacterial and fungal contamination.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (18 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (4 mentions)
Mc3t3 e1 preosteoblasts, by RIKEN Cell Bank (3 mentions)
Alpha mem, by Nacalai Tesque (3 mentions)
Fetal bovine serum (fbs), by Nacalai Tesque (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Dulbecco s modified eagle s medium, by Nacalai Tesque (3 mentions)
Dulbecco s modified eagle s medium, by Fujifilm (2 mentions)
Minimal essential medium, by Nacalai Tesque (2 mentions)
Dulbecco modified eagle medium (dmem), by Nacalai Tesque (2 mentions)
Rpmi 1640 medium, by Fujifilm (2 mentions)
Heat inactivated fetal bovine serum, by Biowest (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Antibiotic antimycotic mixed stock solution, by Fujifilm (2 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Penicillin, by Nacalai Tesque (2 mentions)
Streptomycin, by Nacalai Tesque (2 mentions)
Amphotericin b, by Nacalai Tesque (2 mentions)
Fetal bovine serum (fbs), by Biowest (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
45 protocols using antibiotic antimycotic mixed stock solution
1
MC3T3-E1 Preosteoblast Culture Protocol
2
C2C12 Myoblast Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C2C12 myoblast cells were cultured in Dulbecco's modified Eagle's medium (Wako Pure Chemical) supplemented with a 1% antibiotic-antimycotic mixed stock solution (Nacalai Tesque) and 10% fetal bovine serum (MP Biomedicals, Solon, OH).
3
Labeling Shark Embryos with CM-DiI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the injection of shark embryos at stage 25, eggs were removed from the seawater tank and briefly incubated on ice. A small window was opened on the surface of the egg case just above the embryo. Embryos were then anesthetized with 20 μl of a mixed solution of 1% ethyl 3-aminobenzoate methanesulfonate (MS-222) (E10521; Sigma) and 2% sodium carbonate (1:1 volume:volume). CM-DiI (C7001; Thermo Fisher Scientific, USA) was prepared as previously described [29 (link)] and microinjected into HCs by using a microinjector (MN-151; Narishige, Japan). After injection, 200 μl of 0.2% antibiotic-antimycotic mixed stock solution (09366–44; Nacalai Tesque, Inc., Japan) in PBS was added to the surface of the embryo. The eggshell was sealed with a polycarbonate filter (GTBP01300; Merck Millipore, USA) using cyanoacrylate adhesive (Aron Alpha, also known as ‘Krazy Glue’; Toagosei, Japan), to prevent air bubbles from entering the eggshell and to prevent the contents of the egg from protruding through the opening. The injected embryos were left to develop for 6–7 weeks at 16 °C in seawater with 0.2% antibiotic-antimycotic mixed stock solution without aeration. The incubation seawater was replaced multiple times weekly.
4
Rat Femur Bone Marrow Mesenchymal Stem Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experiments were performed in compliance with the National Animal Care Guidelines (approval no. 20–08004, 31 July 2020). Femurs were isolated from two male rats aged eight weeks, clipped at both ends, and flushed with media using a 21-gauge needle to collect the bone marrow. Bone marrow mesenchymal stem cells were then cultured in 75 cm2 flasks according to a well-documented method [31 (link)]. Primary cells were cultured at 37 °C in a 5% CO2 atmosphere in a growth medium containing minimal essential medium (Nacalai Tesque, Inc., Tokyo, Japan) supplemented with 10% fetal bovine serum (Nacalai Tesque, Inc.) and antibiotic–antimycotic mixed stock solution (Nacalai Tesque, Inc.). The media were changed every three days. rBMMSCs were removed from the flasks at the 3rd–5th culture and seeded at a cell density of 4 × 104 cells/well onto the specimen surface.
5
Cultivation of HPV16-positive Cervical Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPV16-positive cervical cancer cell lines CaSki and SiHa (purchased from ATCC, Virginia, USA) were maintained in Dulbecco’s Modified Eagle Medium supplemented with 10% FBS (Life Technologies, California, USA) and antibiotics (Antibiotic-Antimycotic Mixed Stock Solution; Nacalai Tesque, Kyoto Japan). The cells were grown in a humidified tissue culture incubator at 37° C in 5% CO2.
6
HT29 Cell Culture and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Enterocyte-like HT29 cells (39 (link)) were used for analysis with the MUSE Cell Analyzer (Merck KGaA, Darmstadt, Germany). HT29 cells were obtained from American Type Culture Collection (Manassas, VA, USA). Cells were routinely grown in Dulbecco's modified Eagle's minimal essential medium (DMEM) (Nacalai Tesque, Inc., Kyoto, Japan) supplemented with 10% sterilized fetal bovine serum (Valley Biomedical, Inc., Winchester, VA, USA) and 1% antibiotic/antimycotic mixed stock solution (Nacalai Tesque, Inc.). HT29 cells were incubated in 5% CO2 at 37˚C. Cell treatment was performed as previously described (40 (link)).
7
Culturing Human Lung Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human embryonic kidney 293FT cells (Thermo Fisher Scientific) were cultured in high-glucose DMEM (Fujifilm Wako) supplemented with 10% fetal bovine serum (FBS). H3122 cells, which were gifted by JA Engelman (Massachusetts General Hospital Cancer Center, Boston, Massachusetts, USA), were cultured in RPMI-1640 medium (Wako Pure Chemical Industries) supplemented with 10% FBS and 100 μg/mL of kanamycin. HCC78 was obtained from DSMZ (Germany). HCC78xe3 ROS1-WT cell is a subclone of HCC78, generated by repeating subcutaneous implantation and in vitro cell culture 3 times, and induced SLC34A2-ROS1 overexpression as previously described (84 (link)). ALK fusion–positive and ROS1 fusion–positive NSCLC PDC lines were established from the patients’ pleural effusion. All patients provided informed consent for the genetic and cell biological analyses, which were performed in accordance with a protocol approved by the Institutional Review Board of the JFCR. NSCLC PDC lines JFCR-028-3 and JFCR-168 and HCC78 were cultured in RPMI and Ham’s F12 medium with 10 mM HEPES (Nacalai Tesque), 15% FBS, and 1× antibiotic-antimycotic mixed stock solution (Nacalai Tesque).
8
Senescence Studies of Human Lung Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A TIG-3 cell line (ATCC) isolated from human fetal lung fibroblasts was used in the present study. TIG-3 cells are thought to show replicative senescence at a population doubling level (PDL) of 70 or higher [34 (link)]. The cells were cultured in an incubator at 37 °C and 5% CO2 in Dulbecco’s Modification of Eagle’s Medium (DMEM) supplemented with 10% FBS and 1% Antibiotic Antimycotic Mixed Stock Solution (Nacalai Tesque, Kyoto, Japan). Cells were seeded at 2 × 105 cells/2 mL medium/well when the passage was conducted after 3 days, and 1 × 105 cells/2 mL medium/well when the passage was conducted after 4 days. At PDL70 and higher, the passage was conducted once a week, and the seeding density was 1 × 105 cells/2 mL medium/well. Cells whose PDLs are around 40 were used as “early passage group” cells. Cells were treated with the test substance from the start of the culture (cells around PDL40), and the test substance was added regularly until proliferation ceased (“late passage group” cells).
9
Extraction of DNA and RNA from Burkitt's Lymphoma Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Jijoye, a latency type III EBV-positive Burkitt’s lymphoma cell line, and BL2, an EBV-negative Burkitt’s lymphoma cell line, which were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany), were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Tokyo, Japan) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Tokyo, Japan), and 1% Antibiotic-Antimycotic Mixed stock solution (Nacalai Tesque, Kyoto, Japan), and were incubated in 5% CO2 at 37°C.
Total DNA and RNA were extracted from Jijoye and BL2 cells, and from each 10 mL of cultured media, respectively. Total DNA and RNA were extracted using the QIAamp DNA Mini kit (Qiagen, Tokyo, Japan) and RNeasy Plus mini Kit (Qiagen, Tokyo, Japan), according to the manufacturer’s protocol, respectively.
Total DNA and RNA were extracted from Jijoye and BL2 cells, and from each 10 mL of cultured media, respectively. Total DNA and RNA were extracted using the QIAamp DNA Mini kit (Qiagen, Tokyo, Japan) and RNeasy Plus mini Kit (Qiagen, Tokyo, Japan), according to the manufacturer’s protocol, respectively.
10
Isolation and Culture of Mouse Mammary Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse mammary tissue was prepared as previously described [34 (link)]. Briefly, about 2 g of mammary tissue was minced into small pastes (approximately 1 mm cubes). It was centrifuged three times at 250 g for 5 min each time. The small pieces of mammary gland were suspended in four 36 mm culture plates, each containing about 125 mg tissue and 2.5 mL DMEM/F12 containing 15% KnockOut™ serum replacement (Gibco BRL), 1% nucleosides (Millipore Co., Billerica, MA, USA), 1% nonessential amino acids (Gibco BRL), 1 mM sodium pyruvate (Gibco BRL), and 1% antibiotic-antimycotic mixed stock solution (Nacalai Tesque Inc., Kyoto, Japan). They were cultured at 37°C with 5% CO2. Melatonin (5 mg) was dissolved in 100 μL of ethanol as the stock solution. In this study, melatonin and LPS (ALX-581-014-L001; Enzo Life Sciences, Farmingdale, NY, USA) were added into DMEM/F12 culture medium simultaneously to give final concentrations of 10 μg/mL [31 (link), 32 (link)] and 100 ng/mL [30 (link)], respectively. Mammary tissue was cultured for 6 h. Melatonin-untreated tissue containing ethanol as the solvent at the same concentration (0.08%, w/w) used in the tests were used as a negative control.
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!