절연 및 Endomyocardial Bioptic 샘플의 Arrhythmogenic 심장 근육 병 증 환자에서 심장 중간 엽 기질 세포의 특성
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| IMDM | Gibco by Life Technologies | 12440053 | |
| Fetal Bovine Serum (FBS) | Sigma-Aldrich | F2442 | |
| PENICILLIN STREPTOMYCIN | Life Technologies Italia | 15140122 | |
| Collagenase NB4 | Serva | 17454.02 | |
| Basic fibroblast growth factor | Peprotech | 100-18B | |
| L-glutammine | Sigma-Aldrich | G7513 | |
| PBS | Lonza | 17-516F | |
| Tryple select 1X (cell dissociation reagent) | Gibco | 12563029 | |
| EDTA | Sigma-Aldrich | EDS | |
| Trypsin-EDTA solution | Sigma-Aldrich | T6689 | |
| Bovine Serum Albumin | Sigma-Aldrich | A2058 | |
| 3-Isobutyl-1-methylxanthine | Sigma-Aldrich | I5879 | |
| Hydrocortisone | Sigma-Aldrich | H4001 | |
| Indomethacin | Sigma-Aldrich | I7378 | |
| Oil Red O | Sigma-Aldrich | O0625 | |
| Isopropanol | Sigma-Aldrich | I9516 | |
| Paraformaldehyde (PFA) Solution 4% in PBS | D.b.a. Italia S.r.l. | sc-281692 | |
| Dimethyl Sulfoxide (DMSO) | Sigma-Aldrich | 276855 | |
| Antibody CD14-FITC (MφP9) | Becton Dickinson | 345784 | |
| Antibody Integrin β1 (CD29)-PE (clone MAR4) | Becton Dickinson | 561795 | |
| Antibody PECAM-1 (CD31)- APC (clone 9G11) | R&D Systems | FAB3567A | |
| Antibody Hematopoietic progenitor cell antigen (CD34)-APC (clone 581) | Thermo Fisher Scientific | CD34-581-05 | |
| Antibody H-CAM (CD44)-PE (clone G44-26) | Becton Dickinson | 561858 | |
| Antibody L-CA (CD45)-APC (clone HI30) | Thermo Fisher Scientific | MHCD4505-4 | |
| Antibody THY1 (CD90)-FITC (clone 5E10) | Becton Dickinson | 561969 | |
| Antibody Endoglin (CD105)-APC (clone 266) | Becton Dickinson | 562408 | |
| Antibody HLA-DR-FITC (clone L243) | Becton Dickinson | <br/> 347400 | |
| Gima Quick Plus sterilizer | Gima | 35642 | |
| Bench centrifuge Sigma 3-16K | Sigma Centrifuges | 10330 | |
| Mr. Frosty Freezing Container | Thermo Fisher Scientific | 5100-0001 | |
| AxioVert 200M microscope | Zeiss | B 40-080 e 03/01 | |
| FACS Gallios | Beckman Coulter | 773231AF | |
| ImageJ (image processing program) | NIH | ||
| Stericup filter units | Merck S.P.A. | SCGPU05RE | |
| Conical Tubes, 50 mL | Eppendorf | 30122178 | |
| Conical Tubes, 15 mL | Eppendorf | 30122151 | |
| Safe-Lock Tubes, 2 mL | Eppendorf | 30120094 | |
| 100 mm TC-Treated Cell Culture Dish | Corning | 430167 | |
| 60 mm TC-Treated Cell Culture Dish | Corning | 430166 | |
| 6 well TC-Treated Multiple Well Plates | Corning | 3516 | |
| Polystyrene Round-Bottom Tubes, 5 mL | BD Falcon | 352058 | |
| BRAND counting chamber BLAUBRAND Bürker-Türk | Sigma-Aldrich | BR719520 |
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Cite This Article
Pilato, C. A., Stadiotti, I., Maione, A. S., Saverio, V., Catto, V., Tundo, F., Dello Russo, A., Tondo, C., Pompilio, G., Casella, M., Sommariva, E. Isolation and Characterization of Cardiac Mesenchymal Stromal Cells from Endomyocardial Bioptic Samples of Arrhythmogenic Cardiomyopathy Patients. J. Vis. Exp. (132), e57263, doi:10.3791/57263 (2018).