Human fibroblasts from three apparently healthy individuals (GM00038, GM08680, GM05756) were purchased from Coriell. Reprogramming was performed using the CytoTune-iPS 2.0 Sendai reprogramming kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Briefly, fibroblasts were plated at 20,000 cells/cm2 (link) and cultured until they were 30–60% confluent. Cells were transduced with the Sendai reprogramming vectors at an MOI of 5:5:3 (KOS:hc-Myc:hKlf4). Cells were maintained in fibroblast media until passaged onto VN-coated plates on day 7. On day 8, medium was changed to E8 Medium and cells were cultured for additional 20 days. On day 28, colonies were picked using a 25-gauge needle to cut single colonies into 6–8 pieces and transferred onto VN-coated plates containing E8 Medium only or supplemented with Y-27632 or CEPT for 48 h. After 48 h, cells were kept in E8 Medium only. Confluency was assessed 8 days after colony picking using the Celigo Imaging Cytometer (Nexcelom Biosciences).
Gm05756
Manufactured by Coriell Institute for Medical Research
Sourced in United Kingdom
GM05756 is a cell line derived from human skin fibroblasts. This cell line is available for purchase from the Coriell Institute for Medical Research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Glutamax, by Thermo Fisher Scientific (4 mentions)
Gm00038, by Coriell Institute for Medical Research (2 mentions)
Gm08680, by Coriell Institute for Medical Research (2 mentions)
Cytotune ips 2.0 sendai reprogramming kit, by Thermo Fisher Scientific (2 mentions)
Gentamicin, by Thermo Fisher Scientific (2 mentions)
Gm12878, by Coriell Institute for Medical Research (2 mentions)
Hek293, by American Type Culture Collection (2 mentions)
Gm12878, by Thermo Fisher Scientific (2 mentions)
Nih3t3, by American Type Culture Collection (2 mentions)
Hek293, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Gm05659, by Coriell Institute for Medical Research (2 mentions)
Gm08398, by Coriell Institute for Medical Research (2 mentions)
Gm02036, by Coriell Institute for Medical Research (2 mentions)
7 protocols using gm05756
1
Fibroblast Reprogramming to iPSCs
2
Fibroblast Reprogramming to iPSCs
3
Cell Culture Maintenance Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HapMap LCLs and primary fibroblasts (GM05756, GM17101, GM17151, GM17152, GM17153, GM17169, GM17347, and GM17379) were purchased from the Coriell Institute (Camden, NJ). THP-1 cells were purchased from the American Type Culture Collection (Manassas, VA). Cells were maintained at 37°C in a 5% CO2 atmosphere. LCLs and THP-1s were grown in RPMI 1640 medium (Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS) or 15% for the carboplatin screen, 2 mM glutamine, 100 U/ml penicillin-G, and 100 mg/ml streptomycin. Primary fibroblasts were grown in DMEM 15% FBS, 1 mM pyruvate, 1× nonessential amino acids, 2 mM glutamine, 100 U/ml penicillin-G, and 100 mg/ml streptomycin. BMMs were prepared from the femurs of C57BL/6 mice and casp1−/− (Kuida et al., 1995 (link)) mice by culture with 85% supplemented RPMI + 15% L929-cell supernatant. HEK-293T and HeLa cells were grown in DMEM supplemented with 10% FBS, 1 mM glutamine, 100 U/ml penicillin-G, and 100 mg/ml streptomycin. Where indicated, THP-1s were differentiated using 20 ng/ml PMA for 4 h followed by 24 h in the absence of PMA to allow for adhesion and differentiation before assay.
4
Cell Culture Protocols for Various Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue culture cell lines (GM12878, Coriell; NIH/3T3, ATCC CRL-1658; HEK293, ATCC CRL-1554; Primary Fibro., inguinal fibroblast, GM05756, Coriell, passage 7) were cultured in 5% CO2 at 37°C. GM12878 cells were grown in Roswell Park Memorial Institute media (RPMI, Gibco, Cat. 11875093) supplemented with 15% (v/v) fetal bovine serum (FBS, Gibco, Cat. 10082147), 1X L-glutamine (Gibco, Cat. 25030081), 1X Penicillin-Streptomycin (Gibco, Cat. 15140122), and gentamicin (Gibco, Cat. 15750060). HEK293 cells were grown in Dulbecco’s Modified Eagle’s media (DMEM, Gibco, Cat. 11995065), supplemented with 10% FBS (v/v), and 1X L-glutamine. NIH/3T3 cells were grown in the same preparation of DMEM as HEK293 cells. Primary Fibroblasts were cultured in a growth medium comprised of DMEM/F12 (with GlutaMax; Thermo Fisher), 10% fetal bovine serum (FBS; Thermo Fisher, v/v), 1% MEM Non-Essential Amino Acids (Thermo Fisher, v/v), and 1X Penicillin-Streptomycin (Gibco). Adherent cell lines were grown to ~90% confluency at the time of harvest.
5
Generating Human iPSCs from Skin Fibroblasts
6
Generating Human iPSCs from Skin Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Skin fibroblasts derived from healthy donors (GM02036 from 11-year female, GM05659 from 1-year male, GM05756 from 2-month male, and GM08398 from 8-year male) were obtained from Coriell Institute for Medical Research (Camden, NJ). Fibroblasts were cultured in DMEM (Life Technologies 11195–073) supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin and 100 μg/ml streptomycin (Life technologies 10378–016) in a humidified incubator with 5% CO2 and 20% O2 at 37 °C. Cells were routinely trypsinized (0.05% trypsin/EDTA) and passaged until reprogramming. Human iPSCs were maintained on Matrigel (Corning, #354230) in the chemically-defined Essential 8 medium (Life Technologies, #A1517001) and passaged using 0.5 mM EDTA/DPBS, as described previously (Beers et al., 2012 (link)) (Table 1 ).
7
Cell Culture Protocols for Various Cell Lines
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!