Studiare cancro del pancreas staminali caratteristiche cellulari per lo sviluppo di nuove strategie di trattamento
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| Counting Chamber/Hemocytometer | Hausser Scientific Co | 3200 | |
| CASY Cell Counter and Analyzer for proliferation and viability measurement | Roche Innovatis | AG CASY Model TTC 45,60,150 μm | |
| GentleMACS Dissociator | Miltenyi Co | 130-093-235 | |
| GentleMACS C Tubes | Miltenyi Co | 130-093-237 | |
| 24-well Ultra-low Attachment Plates | Corning | 3474 | |
| 100-mm tissue culture dishes | BD Falcon | 353803 | |
| Cell strainer | BD Falcon | 352350 | |
| 15-ml polypropylene conical tube | BD Falcon | 352097 | |
| 50-ml polypropylene conical tube | BD Falcon | 352070 | |
| 1.5 ml sterile tubes | Eppendorf | 0030 120.086 | |
| 50 ml centrifuge tubes | Corning | 430828 | |
| Sterile petri dishes, 10 cm dishes | Corning | 353003 | |
| 26 G needles | BD Falcon | 300600 | |
| 100 ul Hamilton Syringe | Hamilton Syringe Co | 81075 | |
| Collagenase type IV | Stem Cell Technologies | 7909 | |
| RPMI Medium 1640 | Life technologies | 11875-085 | |
| Penicillin/Streptomycin solution, 100X | Life technologies | SV30010 | |
| Metformin | Sigma | D150959-5G | |
| L-glutamine, 200 mM, 100X | Life technologies | 25030-081 | |
| D-Glucose | Sigma | G8270 | |
| 100mM Sodium Pyruvate solution | Life technologies | 11360-070 | |
| Seahorse Assay medium | Seahorse Bioscience | 100965-000 | |
| BD Cell-TAK Cell and Tissue adhesive | BD Biosciences | 354240 | |
| Trypsin solution, 0.05% | Life technologies | 25300054 | |
| B27 Supplement 50x | Life technologies | 17504-044 | |
| Basic Fibroblast Growth Factor | Sigma | F0291 | |
| Bovine Serum Albumin | Sigma | A9576 | |
| Dulbecco’s Modified Eagle Medium/F12 | Sigma | D8437 | |
| Sterile 1x Dulbecco’s Phosphate Buffered Saline | Sigma | D8537 | |
| Trypan Blue | Life technologies | 15250-061 | |
| Carboxy-DCFDA(5-(and-6)-Carboxy-2',7'-DichlorofluoresceinDiacetate) | Life technologies | C-369 | |
| Rotenone | Sigma | R8875 | |
| Ethanol 70% (vol/vol) | Sigma | 459844 | |
| Isofluorane | IsoVet, Braun | 571105.8 | |
| Matrigel | BD Biosciences | 35620 | |
| Sterile Cotton tipped applicator | Puritan medical | ||
| XF96<sup>e</sup> FluxPak with cell plates, calibrant and cartridges | Seahorse Bioscience | 102416-100 | |
| XF96<sup>e</sup> Extracellular Flux analyzer | Seahorse Bioscience | ||
| Incubator with CO<sub>2</sub> input | |||
| Micro scissors | |||
| Curved forceps | |||
| Splinter forceps | |||
| Caliper |
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Cite This Article
Lonardo, E., Cioffi, M., Sancho, P., Crusz, S., Heeschen, C. Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies. J. Vis. Exp. (100), e52801, doi:10.3791/52801 (2015).