Blood samples were collected into heparin tubes (TERUMO, Tokyo, Japan). Plasma isolated from blood samples was frozen at ‐80°C. Peripheral blood mononuclear cells (PBMCs) were isolated by density centrifugation using Ficoll-Paque Plus (GE Healthcare BioSciences, Tokyo, Japan). PBMCs were cryopreserved in liquid nitrogen by CELLBANKER (Nippon Zenyaku Kogyo, Tokyo, Japan).
Cellbanker
Manufactured by Nippon Zenyaku Kogyo
Sourced in Japan
CELLBANKER is a cryopreservation medium designed for the long-term storage of cells. It is a sterile, ready-to-use solution that helps maintain cell viability and functionality during the freezing and thawing process.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll paque plus, by GE Healthcare (1 mentions)
Penicillin, by Nacalai Tesque (1 mentions)
Streptomycin, by Nacalai Tesque (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Heat inactivated fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Merck Group (1 mentions)
Y 27632, by Selleck Chemicals (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Pathway anti her 2 neu 4b5, by Roche (1 mentions)
Penicillin streptomycin solution p s, by Thermo Fisher Scientific (1 mentions)
Matrigel basement membrane matrix, by Corning (1 mentions)
Fetal calf serum (fcs), by Biowest (1 mentions)
Cell strainer, by BD (1 mentions)
Human serum, by Merck Group (1 mentions)
Nutrient mixture f 12 dmem f12, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
6 protocols using cellbanker
1
PBMC Isolation and Cryopreservation
2
Cryopreservation of Iliac Bone-Derived Cells
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We cultured iliac bone samples collected during secondary bone grafting, but unused, in a minimum essential medium (a-MEM medium; Life Technologies Corporation, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma–Aldrich MO, USA), antibiotics (100 U/ml penicillin and 100 g/ml streptomycin), and 1 ng/ml bFGF in 25 cm2 flasks (Sumitomo Bakelite Co, Tokyo, Japan) at 37 °C in 5% CO2, with medium exchanges performed twice per week. At subconfluency, we subcultured the cells in 75 cm2 flasks, suspended in serum-containing CELLBANKER™ (Nippon Zenyaku Kogyo Co., Ltd., Fukushima, Japan), and cryopreserved at −80 °C.
Cryopreserved cells were thawed and recultivated in a-MEM medium at 37 °C in 5% CO2. Then, we harvested the cells at subconfluency. We used secondary passage cultures for all samples in all experiments.
Cryopreserved cells were thawed and recultivated in a-MEM medium at 37 °C in 5% CO2. Then, we harvested the cells at subconfluency. We used secondary passage cultures for all samples in all experiments.
3
Cryopreserved Tumor Culture Establishment
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The surgically resected tumor was first stored in CELLBANKER (Nippon Zenyaku Kogyo Co., Ltd., Fukushima, Japan) at −80 °C. The tumor was stored for 1 year and 11 months before culturing. At the time of primary culture, the cryopreserved tumor specimen was thawed and divided into small pieces with sterile scissors. The tumor specimens were cultured in DMEM/F12 (Gibco, Grand Island, NY, USA) supplemented with 5% heat-inactivated fetal bovine serum (Gibco), 100 μg/mL penicillin and 100 μg/mL streptomycin (Nacalai Tesque, Kyoto, Japan), 0.4 µg/mL hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 5 ng/mL EGF (Sigma-Aldrich), 10 ng/mL bFGF (Sigma-Aldrich), 5 µg/mL insulin (Sigma-Aldrich), and 10 µM Y-27632 (Selleck Chemicals, Houston, TX, USA) at 37 °C in a humidified atmosphere with 5% CO2. The cells that developed from the tumor specimens were maintained for more than 6 months under tissue culture conditions and were passaged more than 25 times.
4
Breast Cancer Immunohistochemistry and FISH
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Dulbecco’s modified Eagle medium (DMEM), F-12 nutrient mixture (Ham’s F-12) and penicillin-streptomycin solution (PS) were purchased from Invitrogen (Carlsbad, CA, USA). Antibiotic-antimycotic mixed stock solutions (AAMS, 100×) were purchased from Nacalai Tesque Inc., (Kyoto, Japan). Fetal calf serum (FCS) was purchased from Biowest (Riverside, MO, USA). Cryopreservation reagents, Cell Banker, were purchased from Nippon Zenyaku Kogyo (Fukushima, Japan). Matrigel basement membrane matrix was purchased from Corning (Corning, NY, USA).
PAXgene tissue fix containers were purchased from QIAGEN (Tokyo, Japan). PATHWAY anti-HER-2/neu (4B5), INFORM HER2 Dual ISH DNA Probe Cocktail Assay (780-4422), and EBER 1 DNP Probe (760-1209) were purchased from Roche Diagnostics (Tokyo, Japan)
PAXgene tissue fix containers were purchased from QIAGEN (Tokyo, Japan). PATHWAY anti-HER-2/neu (4B5), INFORM HER2 Dual ISH DNA Probe Cocktail Assay (780-4422), and EBER 1 DNP Probe (760-1209) were purchased from Roche Diagnostics (Tokyo, Japan)
5
Isolation of Human Auricular Chondrocytes
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This study was approved by the Research Ethics Committee of the University of Tokyo Hospital. Auricular cartilages were provided as excised remnant auricular cartilage tissue from the surgery of microtia patients in NAGATA Microtia and Reconstructive Plastic Surgery Clinic. We obtained informed consent from all patients. After the excision of soft tissues and perichondria by scalpel and scissors, auricular cartilage was minced, and digested by shaking with 0.3% collagenase solution for 18 h at 37 °C. The solution was filtered with a cell strainer (100 μm pore size, BD Falcon), centrifuged at 1500 rpm for 5 min and the supernatant was removed to obtain human auricular chondrocytes. Cells were seeded at 2.0 × 105 cells/dish to φ100 mm collagen Type I Coated dich (AGC Techno Glass Co., Ltd.), and cultured in the cartilage growth medium (HFI; Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12; Sigma–Aldrich Co.) supplemented with 5% Human Serum (Sigma–Aldrich Co.), 100 ng/mL FGF-2 (Kaken Pharmaceutical Co, Ltd.), 5 μg/mL Insulin (Novo Nordisk Pharma Ltd.), 1% Penicillin/Streptomycin (Sigma–Aldrich Co.)) at 37°C in 5% CO2. After 10 days, cells reached the confluence, and they were detached by Trypsin-EDTA (Sigma–Aldrich Co.), recovered by centrifugation, and stored frozen at −80°C in CELLBANKER (Nippon Zenyaku Kogyo Co., Ltd.).
6
AML cells and CD34+ CB cells
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BM cells were collected from AML patients, which contained >70% AML cells. Bone marrow mononuclear cells (BMMNCs) were isolated by Ficoll centrifugation and cryopreserved in Cellbanker (Nippon Zenyaku Kogyo Co, Fukushima, Japan) until use. hCD34+ CB cells from a single donor were purchased from Promo Cell (Heidelberg, Germany). Freshly thawed cells were used in all experiments.
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