The dental papilla was separated from maxillary and mandibular molar tooth germ of neonatal mouse and digested in a solution containing type I collagenase (3 mg/ml) (catalog no. C0130; Sigma-Aldrich, St. Louis, MO) for 25 min at 37°C. Single-cell suspensions and dental papilla tissue were cultured in EpiCM (epithelial cell medium) basal medium supplemented with 2% fetal bovine serum (FBS) (catalog no. 0010, STEMCELL Technologies, USA), 1% cell growth supplement [EpiCGS (epithelial cell growth supplement), catalog no. 4152, STEMCELL Technologies, USA], and 1% penicillin/streptomycin solution (catalog no. 0503, STEMCELL Technologies, USA). DPCs P2 were used for this study. All cells were grown in a humidified incubator (Thermo Fisher Scientific, USA) at 37°C with 5% CO2, and the medium was refreshed every 3 days.
Penicillin streptomycin solution
Manufactured by STEMCELL
Sourced in United States
Penicillin/streptomycin solution is a sterile, cell culture-tested liquid containing penicillin and streptomycin antibiotics. It is commonly used to prevent bacterial contamination in cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Hepes, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Humidified incubator, by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Immunocult xf t cell expansion medium, by STEMCELL (1 mentions)
Human ab serum, by Merck Group (1 mentions)
Aim 5 medium, by Thermo Fisher Scientific (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Penicillin streptomycin solution, by Merck Group (1 mentions)
Texmacs medium, by Miltenyi Biotec (1 mentions)
Human igf 2, by Merck Group (1 mentions)
A83 01, by STEMCELL (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
3 protocols using penicillin streptomycin solution
1
Isolation and Culture of Mouse Dental Papilla Cells
2
Comparative Analysis of T-cell Culture Media
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Five culture media were used in this study. (1) Complete CTL:AIM-V Medium, used in REP, consisted of equal parts of supplemented RPMI 1640 and AIM-V Medium. To a 1× RPMI 1640 Medium basal, the following components were added: 2 mM L-Glutamine (Fisher Scientific), 10% heat-inactivated (56°C, 60 min) human AB serum (Sigma), 12.5 mM HEPES (Fisher), 1× penicillin streptomycin solution (Fisher), and 50 μM β-mercaptoethanol (Sigma). AIM-V Medium (Invitrogen) was supplemented with 20 mM HEPES and 2 mM L-Glutamine (CTL:AIM-V). (2) ImmunoCult-XF T cell Expansion Medium (STEMCELL Technologies) was supplemented with 1× penicillin streptomycin solution (ICM). (3) TexMACS medium (Miltenyi) was supplemented with 3% heat-inactivated human AB serum (Sigma) and 1× penicillin streptomycin solution (TAC). (4) Corning medium consisted of Lymphocyte Serum-Free Medium KBM 581 (Corning) supplemented with 3% heat-inactivated human AB serum (CMM). (5) Basal HPLM (kindly provided by Dr. Jason Cantor)21 (link) was prepared with four additional components (5 μM acetylcarnitine, 5 μM α-ketoglutarate, 5 μM malate, and 3 μM uridine), as recently reported,51 and then further supplemented with 3% heat-inactivated human AB serum (HPLM). All complete media were filtered through a 0.22-μM filter prior to use.
3
2D and 3D Colony Formation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the 2D colony formation assay, 500 cells were cultivated in 6-well plates for 7-14 days. Colonies were stained with crystal violet (0.5% dissolved in 25% methanol). The number of colonies was counted using Fiji (Fiji, RRID:SCR_002285) as described (59, 60) .
For the 3D colony formation assay, 500 cells were cultured in 2 mL CnT-Prime Airway Epithelial Proliferation Medium (Cat. #CnT-PR-A; CELLNTEC) supplemented with 0.5% methylcellulose (Cat. #M0262; Sigma-Aldrich); 10ng/mL Human IGF-II (Cat. #100-12; PeproTech); 10ng/mL Human Heregulinβ-1 (Cat. #AF-100-03; PeproTech); 1 µM DMH-1 (Cat. #73632; STEMCELL Technologies); 1 µM A-83-01 (Cat. #72022; STEMCELL Technologies); 1% penicillin/streptomycin solution (Cat. #P0781, Sigma-Aldrich). in Nunclon Sphera 6-Well Plate (Cat. #174932; Thermo Fisher Scienti c). After 2-3 weeks, spheres were counted under the microscope, and images were taken at 40X magni cation.
For the 3D colony formation assay, 500 cells were cultured in 2 mL CnT-Prime Airway Epithelial Proliferation Medium (Cat. #CnT-PR-A; CELLNTEC) supplemented with 0.5% methylcellulose (Cat. #M0262; Sigma-Aldrich); 10ng/mL Human IGF-II (Cat. #100-12; PeproTech); 10ng/mL Human Heregulinβ-1 (Cat. #AF-100-03; PeproTech); 1 µM DMH-1 (Cat. #73632; STEMCELL Technologies); 1 µM A-83-01 (Cat. #72022; STEMCELL Technologies); 1% penicillin/streptomycin solution (Cat. #P0781, Sigma-Aldrich). in Nunclon Sphera 6-Well Plate (Cat. #174932; Thermo Fisher Scienti c). After 2-3 weeks, spheres were counted under the microscope, and images were taken at 40X magni cation.
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!