U-2 OS

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Osteosarcome (originally called 2T) derived from a moderately differentiated sarcoma of the tibia of a 15 year old human female (Ponten and Saksela (1967) Int. J. Cancer 2: 434-447).

Cell Type:
Bone
Tissue Origin:
bone
Species:
human
Research Area:
Cancer Research/Cell Biology
Cell Characteristics:
Adherent

Recommended Media

McCoy's 5A Medium

McCoy's 5A Medium was developed in 1956 to be formulated for the nutritional requirements of Walker 256 carcinoma. Its unique components include asparagine, 3X normal glucose, amino acids (all Lisomers) and increased folic acid. It is a general purpose medium for primary and established cell lines.

12-168 With L-glutamine and 25 mM HEPES 
12-688 With L-glutamine

Storage = 2ºC to 8ºC

ProCHO

ProCHO™ Protein-free CHO Media were developed specifically to facilitate the production and downstream processing of recombinant proteins expressed in CHO cells. These protein-free formulations support high-density cultures without the need for animal derived components.
Very low levels of recombinant insulin facilitate both downstream purification and regulatory compliance.

The following media systems are available:
- ProCHO™ 4 Medium – For concurrent transition of adherent CHO cells to serum-free and suspension culture; supports faster doubling times
- ProCHO™ 5 Medium – For CHO cells already growing in suspension; supports increased protein production

Storage = 2°C to 8°C

12-029Q - ProCHO™ 4 with phenol red and 0.1% Pluronic® F-68; but without L-glutamine, hypoxanthine, or thymidine.
04-919Q - ProCHO™ 4 with 0.1% Pluronic® F-68; and without phenol red, L-glutamine, hypoxanthine, or thymidine.

12-766Q - ProCHO™ 5  with 0.1% Pluronic® F-68; and without L-glutamine, phenol red, hypoxanthine, or thymidine.
BE15-766 - ProCHO™ 5 powder without phenol red

UltraMDCK

UltraMDCK Serum-free Renal Cell Medium is designed to support the growth of MADIN-DARBY Canine Kidney (MDCK) cells at low and high plating densities.
The medium contains only two proteins: recombinant human insulin and bovine transferrin, yielding a very low protein formulation.

Storage = 2ºC to 8ºC

Transfection Information

Lonza Optimized Protocol
Optimization Guideline
Filter:
The table below shows data for the cell type and Nucleofector™ Platform selected. Those data are either based on Lonza Optimized Protocols or on results shared from customers who performed an optimization based on our guidelines. In case no data are shown for the selected Nucleofector™ Platform, please take a look at our optimization strategy to get further guidance on how to easily determine optimal Nucleofection conditions yourself.
Protocol Kit Program Cells Efficiency Viable Cells Substrate Format Platform
SE CM-104 2e5 99% 91-99% Plasmid (general) 0.4 µg 20 µl 4D X-Unit
SE CM-104 1e6 99% 85-87% Plasmid (general) 2 µg 100 µl 4D X-Unit
V X-001 1e6 97-99% 80-96% Plasmid (general) 2 µg 100 µl I/II/2b

Citations (20)

Efficient C• G-to-G• C base editors developed using CRISPRi screens, target-library analysis, and machine learning
Categories:
Transfection 
Authors:
Koblan LW, Arbab M, Shen MW, Hussmann JA, Anzalone AV, Doman JL, Newby GA, Yang D, Mok B, Replogle JM, Xu A, Sisley TA, Weissman JS, Adamson B, Liu DR. 
In:
Nat Biotechnol (2021) 39(11): 1414-1425 
CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells
Categories:
Transfection 
Authors:
Kurt IC, Zhou R, Iyer S, Garcia SP, Miller BR, Langner LM, Grünewald J, Joung JK 
In:
Nat Biotechnol (2021) Jan;39(1): 41-46 
A bacterial cytidine deaminase toxin enables CRISPR-free mitochondrial base editing 
Categories:
Transfection 
Authors:
Mok BY, de Moraes MH, Zeng J, Bosch DE, Kotrys AV, Raguram A, Hsu F, Radey MC, Peterson SB, Mootha VK, Mougous JD, Liu DR 
In:
Nature (2020) 7817: 631-637 
Search-and-replace genome editing without double-strand breaks or donor DNA.
Categories:
Primary Cells and Media, Transfection, Cell Lines and Primary Cancer Cells 
Authors:
Anzalone AV1,2,3, Randolph PB1,2,3, Davis JR1,2,3, Sousa AA1,2,3, Koblan LW1,2,3, Levy JM1,2,3, Chen PJ1,2,3, Wilson C1,2,3, Newby GA1,2,3, Raguram A1,2,3, Liu DR4,5,6. 
In:
Nature (2019) 10: 1038 
Highly efficient editing of the ß-globin gene in patient-derived hematopoietic stem and progenitor cells to treat sickle cell disease
Categories:
Transfection 
Authors:
So Hyun Park , Ciaran M Lee , Daniel P Dever , Timothy H Davis , Joab Camarena , Waracharee Srifa , Yankai Zhang , Alireza Paikari , Alicia K Chang , Matthew H Porteus , Vivien A Sheehan , Gang Bao  
In:
Nucleic Acids Res (2019) 47(15): 7955-7972 
Gene correction for SCID-X1 in long-term hematopoietic stem cells.
Categories:
Primary Cells and Media, Serum-free and Speciality Media, Transfection 
Authors:
Pavel-Dinu M, Wiebking V, Dejene BT, Srifa W, Mantri S, Nicolas CE, Lee C, Bao G, Kildebeck EJ, Punjya N, Sindhu C, Inlay MA, Saxena N, DeRavin SS, Malech H, Roncarolo MG, Weinberg KI, Porteus MH 
In:
Nat Commun. (2019) 10 (1): 1634 
A High-Throughput Platform to Identify Small-Molecule Inhibitors of CRISPR-Cas9
Categories:
Transfection 
Authors:
Maji B, Gangopadhyay SA, Lee M, Shi M, Wu P, Heler R, Mok B, Lim D, Siriwardena SU, Paul B, Dancík V, Vetere A, Mesleh MF, Marraffini LA, Liu DR, Clemons PA, Wagner BK, Choudhary A 
In:
Cell Res (2019) 177(4): 1067-1079 
Marker-free coselection for CRISPR-driven genome editing in human cells.
Categories:
Transfection 
Authors:
Agudelo D, Duringer A, Bozoyan L, Huard CC, Carter S, Loehr J, Synodinou D, Drouin M, Salsman J, Dellaire G, Laganière J, Doyon Y. 
In:
Nat Methods (2017) 14(6): 615-620 
Characterization of the interplay between DNA repair and CRISPR/Cas9-induced DNA lesions at an endogenous locus.
Categories:
Transfection 
Authors:
Bothmer A, Phadke T, Barrera LA, Margulies CM, Lee CS, Buquicchio F, Moss S, Abdulkerim HS, Selleck W, Jayaram H, Myer VE, Cotta-Ramusino C. 
In:
Nat Commun. (2017) 8: 13905 
High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects
Categories:
Transfection 
Authors:
Kleinstiver BP, Pattanayak V, Prew MS, Tsai SQ, Nguyen NT, Zheng Z, Joung JK 
In:
Nature (2016) 529 (7587): 490-5 
Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing.
Categories:
Transfection 
Authors:
Shengdar Q. Tsai, Nicolas Wyvekens, Cyd Khayter, Jennifer A. Foden, Vishal Thapar, Deepak Reyon, Mathew J. Goodwin, Martin J. Aryee, J. Keith Joung 
In:
Nat Biotechnol (2014) 32(6): 569-76 
Improving CRISPR-Cas nuclease specificity using truncated guide RNAs
Categories:
Transfection 
Authors:
Yanfang Fu, Jeffry D. Sander, Deepak Reyon, Vincent M. Cascio, J. Keith Joung 
In:
Nat Biotechnol (2014) 32(3): 279-84 
Identification of Putative Target Genes of the Transcription Factor RUNX2
Categories:
Transfection 
Authors:
M Kuhlwilm, A Davierwala, S Pääbo 
In:
PLoS ONE (2013) 8 (12): e83218 
High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells
Categories:
Transfection 
Authors:
Yanfang Fu, Jennifer A Foden, Cyd Khayter, Morgan L Maeder, Deepak Reyon, J Keith Joung, Jeffry D Sander 
In:
Nat Biotechnol (2013) 31(9): 822-6 
FLASH assembly of TALENs for high-throughput genome editing.
Categories:
Transfection 
Authors:
Reyon D, Tsai SQ, Khayter C, Foden JA, Sander JD, Joung JK. 
In:
Nat Biotechnol (2012) 30(5): 460-465 
Insulin-like growth factor binding protein-5 interacts with the vitamin d receptor and modulates the vitamin d response in osteoblasts
Categories:
Transfection 
Authors:
Schedlich LJ, Muthukaruppan A, O'han MK, Baxter RC 
In:
Mol Endocrinol (2007) 21(10): 2378-90 
A Dominant-Negative Form of the Major Human Abasic Endonuclease Enhances Cellular Sensitivity to Laboratory and Clinical DNA-Damaging Agents
Categories:
Transfection 
Authors:
McNeill DR, Wilson DM 3rd 
In:
Mol Cancer Res (2007) 5(1): 61 - 70 
CHK1 phosphorylates CDC25B during the cell cycle in the absence of DNA damage
Categories:
Transfection 
Authors:
Schmitt E, Boutros R, Froment C, Monsarrat B, Ducommun B, Dozier C 
In:
J Cell Sci (2006) 119(Pt 20): 4269-75 
Requirement of clathrin heavy chain for p53-mediated transcription
Categories:
Transfection 
Authors:
Enari M, Ohmori K, Kitabayashi I, Taya Y 
In:
Genes Dev (2006) 20(9): 1087-99 
Immediate transfection of patient-derived leukemia: a novel source for generating cell-based vaccines
Categories:
Transfection 
Authors:
Gershan JA, Johnson BD, Weber J, Schauer DW, Natalia N, Behnke S, Burns K, Maloney KW, Warwick AB and Orentas RJ 
In:
Genet Vaccines Ther (2005) 3(1): 4 
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