To assemble cell sheets (CS), MSC were seeded at a high density ≈ 15,000 cells/cm2 (60,000 cells per well in a 12-well CELLSTAR culture plate (Greiner Bio-One GmbH, Kremsmünster, Austria) or 300,000 cells per 60-mm Petri dish (Corning, Corning, NY, USA). During the first 7 days, the cell cultures achieved sufficient density for the assembly of CS, and the formation of condensed and scattered areas within CS occurred by days 10–14; throughout CS assembly, the medium was changed every 2 days. A time-lapse observation of the CS assembly was performed in accordance with the methods in Table 1 .
60 mm petri dish
Manufactured by Corning
Sourced in United States
The 60 mm Petri dish is a circular, shallow dish made of glass or plastic, designed for use in laboratories. It serves as a container for various experimental media, samples, or cultures. The dish measures 60 mm in diameter and provides a controlled environment for the growth and observation of microorganisms, cell cultures, or other specimens.
Automatically generated - may contain errors
Lab products found in correlation
Tnf α, by Merck Group (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hela cells, by National Collection of Authenticated Cell Cultures (1 mentions)
Mesenchymal stem cell medium, by ScienCell (1 mentions)
Electronic balance, by Sartorius (1 mentions)
Water bath, by Thermo Fisher Scientific (1 mentions)
Petri dish, by Zeiss (1 mentions)
Stemi 508 trinoc microscope, by Zeiss (1 mentions)
Ultrapure agarose, by Thermo Fisher Scientific (1 mentions)
Illustrator 2020, by Adobe (1 mentions)
Defined trypsin inhibitor, by Thermo Fisher Scientific (1 mentions)
Epilife defined growth supplement, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
6 protocols using 60 mm petri dish
1
Mesenchymal Stem Cell Sheet Assembly
2
Induction of Vascular Smooth Muscle Cell Dysfunction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A rat VSMC cell line A7r5 was purchased from the Bioresource Collection and Research Center. A7r5 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin mixed antibiotics, 1% glutamine, and 1.5 g/L sodium bicarbonate (all regents from Hyclone, Logan, Utah, USA). All cell cultures were maintained at 37 °C under 95% moisturized air with 5% CO2. Before cell treatments, A7r5 cells were seeded onto each 60 mm Petri dish (Corning Inc, Corning, NY, USA) at a density of 105 for 24 h. For induction of VSMC dysfunction, A7r5 cells at 70% confluence were serum-starved for 24 h and treated with TNF-α (10 ng/mL; Sigma-Aldrich, St Louis, MO, USA) for 24 h.
3
HeLa Cell Culture and Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in Minimum Essential Medium (MEM, GIBCO) with 10% fetal bovine serum (FBS, Invitrogen, Life Technologies) in a 5% CO2 incubator at 37 °C. For imaging experiments, cells were seeded on 60 mm Petri dish (Corning) at a density of 300,000 and cultured overnight. Typically, HeLa cells were incubated with 0.1 nM fPlas-gold in MEM medium containing 10% FBS and washed with 1 × PBS buffer (pH 7.4) before imaging. HeLa cell line used in this study is not on the ICLAC and NCBI biosample misidentified cell list. It was not authenticated and not tested for mycoplasma contamination.
4
Isolation of Alveolar BMSCs from Implant Debris
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In this study, human alveolar BMSCs were isolated from wasted bone debris from the implant sockets of patients who underwent oral implantation. An informed consent form for the study was signed by each patient before surgery. The study was approved by the Ethics Committee of Beijing Stomatological Hospital, Capital Medical University (ethics approval: CMUSH-IRB-KJ-PJ-2017-01), and was performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.
The primary BMSC culture method used in this study was similar to that described in our previous study [22 (link)]. During oral implant surgery, the implant sockets were prepared using a low-speed drilling technique (50 rpm, without irrigation), and the bone debris was then collected from the drill and placed in sterile tubes containing 0.5 ml phosphate-buffered saline (PBS) (HyClone, USA). An electronic balance (Sartorius, Germany) was used to weigh the tubes before and after the addition of the bone debris to enable quantification. After centrifugation, the bone debris was transferred into a 60 mm Petri dish (Corning, USA) with 5 ml of mesenchymal stem cell medium (MSCM) (ScienCell, USA) and placed in a 37°C and 5% CO2 incubator for 7 d. Thereafter, the medium was replaced every 3 d.
The primary BMSC culture method used in this study was similar to that described in our previous study [22 (link)]. During oral implant surgery, the implant sockets were prepared using a low-speed drilling technique (50 rpm, without irrigation), and the bone debris was then collected from the drill and placed in sterile tubes containing 0.5 ml phosphate-buffered saline (PBS) (HyClone, USA). An electronic balance (Sartorius, Germany) was used to weigh the tubes before and after the addition of the bone debris to enable quantification. After centrifugation, the bone debris was transferred into a 60 mm Petri dish (Corning, USA) with 5 ml of mesenchymal stem cell medium (MSCM) (ScienCell, USA) and placed in a 37°C and 5% CO2 incubator for 7 d. Thereafter, the medium was replaced every 3 d.
5
Immobilizing and Imaging Adult Fly Heads
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adult flies were immobilized by embedding them in an agarose gel. To prepare the agarose gel, we dissolved 2 g of UltraPure Agarose (Invitrogen, Waltham, MA, USA) in 100 mL distilled water. The solution was heated in a 500 mL Erlenmeyer flask for several minutes until bubbles were visible. The flask was then placed in a water bath (Thermo Scientific, Waltham, MA, USA) set to 58 °C to cool the solution down to 58 °C. Next, flies were anesthetized with carbon dioxide gas and transferred to a 60 mm petri dish (Corning, Corning, NY, USA) filled with about 10 mL of the liquid agarose gel. We then submerged the wings and legs using forceps and placed the petri dish on ice for the gel to solidify. After solidification of the gel, the petri dish was placed under a Stemi 508 Trinoc microscope (model #4350649030, Zeiss, White Plains, NY, USA), and the fly head was adjusted with the forceps such that one eye was oriented in parallel to the microscope lens. The petri dish was placed back on ice until imaging, which was performed with an Axiocam 208 HD/4k color camera (model #4265709000) set to auto exposure and auto white balance. Images were processed with Fiji (https://imagej.net/software/fiji/ accessed on 10 July 2022) [24 (link)], Adobe Photoshop 2020, and Adobe Illustrator 2020 software.
6
Establishing Primary Lip Keratinocyte Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary keratinocyte cultures from three sections of lip tissue were established using the explant culture technique. Small explants (dermal side up) were placed in a 60 mm Petri dish (Corning, Corning, NY, USA). They were incubated in a moist atmosphere at 37 • C and 5% CO 2 with complete EpiLife that contained EpiLife-defined growth supplements (Thermo Fisher Scientific, Waltham, MA, USA), 0.06 mM Ca 2+ , gentamicin (5.0 µg/mL; Thermo Fisher Scientific, Waltham, MA, USA), and amphotericin B (0.375 µg/mL; Thermo Fisher Scientific, Waltham, MA, USA), which is a serum-free culture medium. The culture medium was refreshed with complete EpiLife medium every other day. When cell outgrowth reached 80% confluence, the p0 keratinocytes were detached with 0.025% trypsin and ethylenediaminetetraacetic acid (Thermo Fisher Scientific, Waltham, MA, USA), neutralized with defined trypsin inhibitor (Thermo Fisher Scientific, Waltham, MA, USA), and replated as p1 cells into another tissue-culture Petri dish (1.0 × 10 4 cells/cm 2 ) with complete EpiLife medium. Passage 1 (p1) keratinocytes obtained from the skin, vermilion, and oral mucosa were fed with the same culture medium every other day. After a confluence of approximately 80% was reached, the total RNA was extracted from the cells using a RNeasy mini kit (Qiagen, Valencia, CA, USA), according to the manufacturer's instructions.
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!