Mice were euthanized and whole blood, draining lymph nodes, spleens, femurs, tibias, and femoral muscles at the injection site were obtained. Sera were prepared from the blood samples and stored at −80°C until the assays were performed. The spleens were dissociated into cell suspensions using a gentleMACS Dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany). The red blood cells were lysed with Red Blood Cell Lysis Buffer (Roche, Basel, Switzerland), and the splenocytes and bone marrow cells were suspended in Roswell Park Memorial Institute-1640 medium (Nacalai Tesque Inc., Kyoto, Japan) supplemented with 10% fetal bovine serum (Sigma-Aldrich, Burlington, MA, USA). The draining lymph nodes were collected, and single-cell suspensions were prepared using BiomusherII (Nippi Research Institute of Biomatrix, Ibaraki, Japan). Femoral muscle samples were fixed with 10% neutral buffered formalin (Mildform 10N; FUJIFILM Wako Pure Chemical, Osaka, Japan) for 24 hours at room temperature (RT), paraffin-embedded, sectioned, and stained with hematoxylin and eosin (H&E).
Roswell park memorial institute 1640 medium
Manufactured by Nacalai Tesque
Sourced in Japan
Roswell Park Memorial Institute-1640 medium is a cell culture medium that is commonly used for the in vitro cultivation of a variety of cell types, including human and animal cells. It provides the necessary nutrients, growth factors, and other components to support the growth and maintenance of these cells in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Mildform 10n, by Fujifilm (1 mentions)
Gentlemacs dissociator, by Miltenyi Biotec (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Red blood cell lysis buffer, by Roche (1 mentions)
Dulbecco s modified eagle s medium, by Nacalai Tesque (1 mentions)
Hgc27, by Nacalai Tesque (1 mentions)
Nugc 4, by RIKEN BioResource Center (1 mentions)
2 protocols using roswell park memorial institute 1640 medium
1
Comprehensive Immune Response Analysis
2
Gastric Cancer Cell Lines and Culture
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The human GC cell lines MKN7 (RCB0999), MKN45 (RCB1001), MKN74 (RCB1002), HGC27 (RCB0500), and NUGC4 (RCB1939) were purchased from Riken BioResource Center (Tsukuba, Ibaragi, Japan), and MKN74/ CMV-Luc (JCRB1473), a human GC cell line, which stably expresses luciferase, was purchased from the Japanese Collection of Research Bioresources Cell Bank (Ibaraki, Osaka, Japan). The human normal peritoneal mesothelial cell line MeT-5A (CRL-9444) was purchased from the American Type Culture Collection (Manassas, VA, USA). HGC27 cells were cultured in Dulbecco's modified Eagle's medium (Nacalai Tesque, Kyoto, Japan). All other cells were cultured in Roswell Park Memorial Institute-1640 medium (Nacalai Tesque). All media were supplemented with 10% sEV-free fetal bovine serum (System Biosciences, Palo Alto, CA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin. All cell lines were cultured in a humidified incubator at 37 °C with 5% CO 2 .
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