Human foreskin fibroblasts (CRL2522; ATCC) and human adult low calcium high temperature keratinocytes (HaCat; CLS) were maintained in growth medium containing DMEM (with 3.7 g/l NaHCO3, with 4.5 g/l glucose and stable L-glutamine; Merck), 10% fetal calf serum (FCS; Pan Biotech) and were supplemented with 1 ng/ml basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) (Pan Biotech). The cells were cultured under humidified conditions with 10% CO2.
Crl 2522
Manufactured by American Type Culture Collection
Sourced in United States
The CRL-2522 is a cell line from the American Type Culture Collection (ATCC). It is derived from human embryonic kidney cells. The CRL-2522 cell line is a common tool used in various cell biology research applications.
Automatically generated - may contain errors
Lab products found in correlation
Hb 8065, by American Type Culture Collection (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Ccl 2, by American Type Culture Collection (4 mentions)
Mcf 7, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Htb 22, by American Type Culture Collection (3 mentions)
Venor gem mycoplasma detection kit, by Merck Group (2 mentions)
Gm03529 cells, by Coriell Institute for Medical Research (2 mentions)
Hek293t 17, by American Type Culture Collection (2 mentions)
Wa09 hescs, by WiCell (2 mentions)
Nutristem hpsc xf medium, by WiCell (2 mentions)
Stemflex medium, by Thermo Fisher Scientific (2 mentions)
Mrna reprogramming kit, by American Type Culture Collection (2 mentions)
Dmem f12, by Bio-Techne (2 mentions)
Mtesr1 medium, by STEMCELL (2 mentions)
Crl 10782, by American Type Culture Collection (2 mentions)
Penicillin, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Lenti x concentrator, by Takara Bio (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (2 mentions)
48 protocols using crl 2522
1
Culturing Human Skin Cell Lines
2
Cell Line Authentication and Maintenance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human BJ dermal fibroblasts (CRL-2522) and HEK293T/17 (CRL-11268) cells were purchased from ATCC and maintained in DMEM supplemented with 10% FBS, NEAA, and β-ME. GM03529 cells were obtain from Coriell Institute and maintained in DMEM supplemented 15% FBS, NEAA, and β-ME. WA09 hESCs were from WiCell Institute and maintained in NutriStem® hPSC XF medium. No cell lines used in this study are listed in the International Cell Line Authentication Committee (ICLAC) Register of Misidentified Cell Lines or the NCBI Biosample database of misidentified cell lines. All cell lines were authenticated by Interspecies Determination (Isoenzyme Analysis and STR analysis) by the providing company and were routinely tested for mycoplasma detection using a Venor GeM Mycoplasma Detection Kit (Sigma-Aldrich).
3
Toxicity Assessment of Nanoparticles on BJ Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For biological testing, we used BJ human skin fibroblasts cultures (ATCC™ CRL-2522) which were derived from an original ATCC batch. Fibroblasts were cultured in Eagle's Minimum Essential Medium (EMEM) supplemented with 10% Fetal Bovine Serum (FBS), 100 U/ml penicillin and 100 μg/mL streptomycin. Cells were incubated at 37 °C in a humidified atmosphere of 5% CO2.
For toxicity assays, BJ cells were inoculated at 5 × 104 cells/mL in 96-well plates and incubated for 24 h. Then, the medium of each well was removed and 100 μL of the nanoparticle suspensions in different concentrations in the cell culture medium was added to wells. In untreated cells, only a complete medium was added. The cells were incubated for different exposure times.
For toxicity assays, BJ cells were inoculated at 5 × 104 cells/mL in 96-well plates and incubated for 24 h. Then, the medium of each well was removed and 100 μL of the nanoparticle suspensions in different concentrations in the cell culture medium was added to wells. In untreated cells, only a complete medium was added. The cells were incubated for different exposure times.
4
Skin Cell Culture Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experiments were performed on two human skin cell lines: HaCaT immortalized keratinocytes (CLS Cell Lines Service GmbH, Eppelheim, Germany) and BJ normal fibroblasts (ATCC® CRL2522™, Manassas, USA). HaCaT cells were cultured in DMEM medium supplemented with 4500 mg/l glucose, 200 mM L-glutamine, 10 U/ml penicillin, 100 mg/ml streptomycin and 10% Fetal Bovine Serum. BJ cells were cultured in EMEM medium supplemented with 200 mM L-glutamine, 10 U/ml penicillin, 10 mg/ml streptomycin and 10% Fetal Bovine Serum. The cells were cultured according to the manufacturer’s instructions at 37ºC and 5% CO2. Experiments were performed between 19 and 25 passage of HaCaT cells and 8–16 passage of BJ cells.
5
Generation of LRRK2 and AARS1 hiPS Cells
6
Generating Cerebral Organoids from iPSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Male human fibroblasts (CRL-2522, ATCC) were used to induce pluripotent stem cells (iPSCs). The use of iPS cells to generate cerebral organoids was approved by the Ethics Commission of LMU (Ludwig-Maximilians-Universität München), with the associated number 115-16. IPSCs and human organoids were cultured at 37°C, 5% CO2 and ambient oxygen level. During the whole period of cerebral organoid generation the medium was changed every day. Electroporations were performed in cerebral organoids at 39 days after the initial plating of the cells and fixed 7 days post-electroporation.
7
Chemotaxis Assay for hFDM Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To prepare proper samples for chemotaxis study, hFDM was freeze-dried and digested using 1 mg/mL pepsin in 0.01 N HCl at 37 °C for 48 h. This hFDM suspension was then neutralized via 0.1 N NaOH and the protein content was quantified using a BCA protein assay kit (23250; Thermo Scientific). For chemotaxis effect, three different concentrations (25, 50, and 100 μg/mL) of soluble hFDM were prepared in DMEM without the addition of FBS. For comparison, DMEM with 10% FBS served as a positive control and without serum (0% FBS) was a negative control. Cell migration across transwell inserts was investigated using two types of cells, human umbilical vein endothelial cells (HUVECs) (C2517A; Lonza) and human dermal fibroblasts (HDFs) (CRL-2522; ATCC). Cells were seeded at 5×104 cells in 8 μm pore diameter transwells (353182; Falcon) filled with medium without FBS. The lower chamber was occupied with hFDM suspension, negative control, or positive control. The overall experimental schematic is shown in Figure S1 . Cell migration was allowed for 3 and 6 h with HDFs and HUVECs, respectively. Then, cells in the lower chamber were stained with 0.2% (w/v) crystal violet and counted under a light microscope (Axio Vert.A1; Carl Zeiss). In each well, five random images (3×105 μm2) from three replicates of each group were taken and the number of cells was determined.
8
Cell Culture Conditions for BJ and HEP G2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cell lines were obtained from the American Type Cell Culture collection, ATCC (LGC Standards-ATCC, Teddington, Great Britain). ATCC designations were as follows: BJ, normal adherent human skin fibroblasts, CRL-2522; HEP G2, hepatocellular carcinoma. Cells were cultured according to ATCC’s catalogue instructions. Briefly, cells were cultured at 37 °C in a humidified atmosphere of 95% air and 5% CO2 to confluence in Eagle’s Minimum Essential Medium (EMEM, ATCC), supplemented with 10% v/v FBS (ATCC) and with antibiotic solution (100 IU/mL penicillin, 0.1 mg/mL streptomycin, Gibco Laboratories, Grand Island, NY). For experiments, the cells were at an exponential phase of growth.
9
Aging hPSC-Derived Cardiac Tissue Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Stem cells were derived from the BJ fibroblast line (CRL-2522, ATCC) by the Washington University Genome Engineering and Stem Cell Core. Differentiation to hPSC-CMs was done in monolayer culture by temporal modulation of WNT signaling as previously described (13 (link), 77 (link)). Cells were aged at least 30 days before performing any assays. All assays were conducted with 2 independent clones and cells were derived from at least 2 independent differentiations. Linear human engineered heart tissue strips were generated as previously described (35 (link)). Ring shaped EHTs were prepared as described in (78 (link)). Ring-shaped engineered heart tissues were mounted in an Aurora Scientific Small Intact Muscle Apparatus between a length mover and a force transducer. Details can be found in the Supporting Materials .
10
Comparative Cytotoxicity and Cytoprotection Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All cell lines utilized in this study were initially obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and were maintained in freezing/storage medium containing 90% fetal bovine serum (FBS)/10% DMSO at −150 °C. Four human cancerous cell lines were used in this study, namely the cervical adenocarcinoma (HeLa, ATCC® CCL-2™), colorectal adenocarcinoma (HCT-116, ATCC® CCL-247™), mammary adenocarcinoma (MCF-7, ATCC® HTB-22™) and hepatocellular carcinoma (HepG2, ATCC® HB-8065™). For comparison of cytotoxicity, the noncancerous skin fibroblast BJ-1 (ATCC® CRL-2522™) and epithelial breast MCF-12 (ATCC® CRL-10782™) cell lines were used. Human amniotic epithelial (WISH, (ATCC® CCL-25™) was used for measuring the cytoprotective activity against oxidative stress. Cell lines were cultured in DMEM/high glucose supplemented with 2 mM L-glutamine, 10% FBS and 1% penicillin/streptomycin. Then, sub-confluent cultures (80–90%) were trypsinized (Trypsin 0.05%/0.53 mM EDTA) and spilt depending on the seeding ratio recommended by ATCC [16 (link),17 (link)].
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!