After removal of the epidermis, the remaining pieces of dermis were transferred to fibroblast medium (Gibco, Waltham) and cut into smaller pieces, which were then placed on a cell-culture dish (Greiner Bio-One) (Figure 3(b) ). After 4 h, the dish was filled with fibroblast medium with the pieces staying adherent to the dish. After several days with regular medium changes, the pieces were removed when fibroblast colonies were visible.
Cell culture dish
Manufactured by Greiner
Sourced in Germany
The cell-culture dish is a laboratory equipment used for growing and maintaining cells in a controlled environment. It provides a sterile and nutrient-rich surface for the cells to adhere and proliferate.
Automatically generated - may contain errors
Lab products found in correlation
Glass bottom, by Greiner (1 mentions)
Orca r2 camera, by Hamamatsu Photonics (1 mentions)
U mwu, by Olympus (1 mentions)
U mwig2, by Olympus (1 mentions)
U mwib, by Olympus (1 mentions)
Aquacosmos, by Hamamatsu Photonics (1 mentions)
Bx51 fluorescence microscope, by Olympus (1 mentions)
Dulbecco s modified eagle medium dmem, by Harvard Bioscience (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (1 mentions)
Bovine serum albumin (bsa), by Avantor (1 mentions)
Cd45 monoclonal antibody, by Abcam (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
4 protocols using cell culture dish
1
Dermal Fibroblast Isolation Protocol
2
Fluorescence Microscopy Visualization of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For fluorescence microscopy, proteins were visualized by fusion to GFP, 3 X mRFP or mCherry. The nuclear chromatin region was stained with bisbenzimide H33342 fluorochrome trihydrochroride (Hoechst 33342). Living cells were observed under a fluorescence microscope (model BX51; Olympus, Tokyo, Japan), and images were obtained by a BX51 fluorescence microscope (Olympus) equipped with an ORCA-R2 camera (Hamamatsu Photonics, Hamamatsu, Japan). Filter sets U-MWU (Olympus), U-MWIB (Olympus), and U-MWIG2 (Olympus) were used for Hoechst 33342, GFP and mCherry/3 X mRFP, respectively. Image acquisition and processing were carried out by using Aquacosmos (Hamamatsu Photonics) and Image J (National Institutes of Health, Bethesda, USA) software. Time-lapse observation was performed as follows. Cells were incubated on SSA at 28°C for 16 hours. After conjugation, cells were inoculated to SSA medium on a cell-culture dish with a glass bottom (Greiner Bio-One, Frickenhausen, Germany) and observed under a fluorescence microscope (model IX71; Olympus) as described in Nakamura et al. (2008) [31 (link)]. Images were processed with Image J.
3
Isolation of Marmoset Bone Marrow Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bone marrow of marmoset was isolated post mortem by rupture of the tibia and femur of each animal immediately after death had been confirmed by a veterinarian. The cavity was flushed with a hypodermic needle attached to a syringe. For preventing coagulation and for cell singularization, a heparin phosphate-buffered saline (PBS) mix was utilized (5 IU/ml) and the bone marrow was separated by pipetting thoroughly for 3 minutes. After singularization, the cell suspension was transferred into red cell lysis buffer (NH4Cl 0.15 M, KHCO3 10 mM, ethylenediaminetetraacetic acid (EDTA) 0.1 mM, pH 7.5) for 5 minutes and centrifuged at 200 × g for 10 minutes. The cell pellet was resuspended in the MSC culture medium consisting of Dulbecco’s Modified Eagle Medium (DMEM; Biochrom AG, Berlin, Germany), 15 % fetal calf serum (FCS; Biochrom AG), 1 % penicillin/streptomycin (Invitrogen GmbH, Karlsruhe, Germany), and 50 μM l -ascorbic acid-2-phosphate and plated on a cell culture dish (Greiner Bio-One GmbH, Frickenhausen, Germany).
4
Immunomagnetic Capture and Identification of Circulating Tumor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-7 cells (passage14) were placed in phosphate-buffered saline (PBS) as mock samples. CTCs were captured by quick-response immunomagnetic nanosphere (IMN)-modified anti-epithelial-cell-adhesion-molecule (EpCAM) monoclonal antibody (Wuhan Jiayuan Quantum Dots Corporation, Ltd., Wuhan, China)21 (link). IMNs were added to 7.5 mL of sample, and the mixture was incubated for 5 min at 37 °C. After magnetic separation, the captured cells were fixed with 4% paraformaldehyde (Nanjing Chemical Reagent Co., Ltd., Nanjing, China), permeabilized, blocked with 0.1% Triton-X 100 (Sigma-Aldrich, MO, USA) and 1% bovine serum albumin (BSA) (Amresco, OH, USA), and stained with 30 μg/mL 4',6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich), FITC-labeled anti-keratin-19 (CK19) monoclonal antibody (Abcam, MA, USA), and APC-labelled anti-protein-tyrosine-phosphatase-receptor-type-C (CD45) monoclonal antibody (Abcam). After washing, the captured cells were placed in a cell culture dish (Greiner, Kremsmünster, Austria) for fluorescence microscopy imaging. CTCs were identified as cells that had round to oval morphology, were positive for DAPI and CK19, and negative for CD45. All processes are completed within 2 h.
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!