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Bioengineering

Los andamios de autoinforme para el 3-Dimensional Cultivo Celular

Published: November 7, 2013 doi: 10.3791/50608

Materials

Name Company Catalog Number Comments
Ethanol Fisher 32221
Anhydrous dimethylformamide (DMF) Sigma 270547
Ammonium hydroxide 50% (v/v) aqueous solution Alfa Aesar 35574 diluted to 30% (v/v) with pure water
TEOS Sigma 13190-3
3-Aminopropyltriethoxysilane (APTES) Sigma A3648
5-(and-6)-carboxyfluorescein, succinimidyl ester (FAM-SE) Invitrogen C1311
6-carboxytetramethylrhodamine, succinimidyl ester (TAMRA-SE) Invitrogen C1171
Sodium phosphate monobasic (0.2 M) Sigma Aldrich S-9638
Sodium phosphate dibasic (0.2 M) Sigma Aldrich S-0876
NaOH Sigma Aldrich S8045
Trypsin/EDTA Sigma Aldrich T4174
Penicillin/Streptomycin Sigma Aldrich P0781
PBS Sigma Aldrich D8537
DMEM Sigma Aldrich D6046
FBS Source Bioscience Batch-213-101992
L-Glutamine Sigma Aldrich G7513
Lipofectamine 2000 Invitrogen 11668-019
Optimem Invitrogen 11058-021
LysoTracker Red Invitrogen L-7528
Draq5 Biostatus Ltd DR50050
Nitrogen gas BOC
DCM Sigma Aldrich 320269
TFA Sigma Aldrich T6508
Confocal microscope Leica TCS-SP equipped with argon and krypton lasers and a 63X 0.9NA water immersion lens
UV light UVLS28 UVP, USA
Stirrer plate SB161-3 Jencons-PLS
pH meter Jenway model 3510
Rotary Evaporator Buchi Rotary Evaporator R200
Centrifuge (nanosensors) Hermle Z300
Centrifuge (cell culture) Thermo Scientific Heraeus Biofuge Primo
Vortex Whirlimixer Fisherbrand
Ultrasonicator FB11021 Fisherbrand
Aluminum sheet Nottingham University
35mm cell culture plate Iwaki 3000035
10 ml syringe Becton Dickenson
3T3 Fibroblast cells European Collection of Cell Cultures
PLGA Lakeshore Biomaterials 7525 DLG 7E
Pyridinium formate Sigma Aldrich P8535
Trypan blue Sigma Aldrich T8154
Sodium phosphate monobasic Sigma Aldrich S9638
Sodium phosphate dibasic Sigma Aldrich S5136
HCl Sigma Aldrich 320331

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References

  1. Takimoto, Y., Dixit, V., Arthur, M., Gitnick, G. De novo liver tissue formation in rats using a novel collagen-polypropylene scaffold. Cell Transplantation. 12 (4), 413-421 (2003).
  2. Sales, V. L., Engelmayr, G. C., et al. Protein precoating of elastomeric tissue-engineering scaffolds increased cellularity, enhanced extracellular matrix protein production, and differentially regulated the phenotypes of circulating endothelial progenitor cells. Circulation. 116 (11), I55-I63 (2007).
  3. Hollister, S. J. Porous scaffold design for tissue engineering. Nature Materials. 4 (7), 518-524 (2005).
  4. Li, D., Xia, Y. N. Electrospinning of nanofibers: Reinventing the wheel? Advanced Materials. 16 (14), 1151-1170 (2004).
  5. Sill, T. J., von Recum, H. A. Electro spinning: Applications in drug delivery and tissue engineering. Biomaterials. 29 (13), 1989-2006 (2008).
  6. Pham, Q. P., Sharma, U., Mikos, A. G. Electrospinning of polymeric nanofibers for tissue engineering applications: A review. Tissue Engineering. 12 (5), 1197-1211 (2006).
  7. Sawyer, N. B. E., Worrall, L. K., et al. In situ monitoring of 3D in vitro cell aggregation using an optical imaging system. Biotechnology and Bioengineering. 100 (1), 159-167 (2008).
  8. Dan, L., Chua, C. K., Leong, K. F. Fibroblast Response to Interstitial Flow: A State-of-the-Art Review. Biotechnology and Bioengineering. 107 (1), 1-10 (2010).
  9. Pancrazio, J. J., Wang, F., Kelley, C. A. Enabling tools for tissue engineering. Biosensors & Bioelectronics. 22 (12), 2803-2811 (2007).
  10. You, Y., Lee, S. W., Youk, J. H., Min, B. M., Lee, S. J., Park, W. H. In vitro degradation behaviour of non-porous ultra-fine poly(glycolic acid)/poly(L-lactic acid) fibres and porous ultra-fine poly(glycolic acid) fibres. Polymer Degradation and Stability. 90 (3), 441-448 (2005).
  11. Dong, Y. X., Liao, S., Ramakrishna, S., Chan, C. K. Distinctive degradation behaviors of electrospun PGA, PLGA and P(LLA-CL) nanofibers cultured with/without cell culture. Multi-Functional Materials and Structures. 47 - 50, 1327-1330 (2008).
  12. Xu, Y. H., Sun, J. J., Mathew, G., Jeevarajan, A. S., Anderson, M. M. Continuous glucose monitoring and control in a rotating wall perfused bioreactor. Biotechnology and Bioengineering. 87 (4), 473-477 (2004).
  13. Harrington, H. C., Rose, F. R. A. J., Reinwald, Y., Buttery, L. D. K., Ghaemmaghami, A. M., Aylott, J. W. Electrospun PLGA fibre sheets incorporating fluorescent nanosensors: self-reporting scaffolds for application in tissue engineering. Analytical Methods. 5 (1), (2013).
  14. Wang, X. Y., Drew, C., Lee, S. H., Senecal, K. J., Kumar, J., Sarnuelson, L. A. Electrospun nanofibrous membranes for highly sensitive optical sensors. Nano Letters. 2 (11), 1273-1275 (2002).
  15. Yang, Y., Yiu, H. H. P., El Haj, A. J. On-line fluorescent monitoring of the degradation of polymeric scaffolds for tissue engineering. Analyst. 130 (11), 1502-1506 (2005).
  16. Blackwood, K. A., McKean, R., et al. Development of biodegradable electrospun scaffolds for dermal replacement. Biomaterials. 29 (21), 3091-3104 (2008).
  17. Bashur, C. A., Dahlgren, L. A., Goldstein, A. S. Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly(D,L-lactic-co-glycolic acid) meshes. Biomaterials. 27 (33), 5681-5688 (2006).
  18. Li, W. J., Laurencin, C. T., Caterson, E. J., Tuan, R. S., Ko, F. K. Electrospun nanofibrous structure: A novel scaffold for tissue engineering. Journal of Biomedical Materials Research. 60 (4), 613-621 (2002).
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Los andamios de autoinforme para el 3-Dimensional Cultivo Celular
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Cite this Article

Harrington, H., Rose, F. R. A. J.,More

Harrington, H., Rose, F. R. A. J., Aylott, J. W., Ghaemmaghami, A. M. Self-reporting Scaffolds for 3-Dimensional Cell Culture. J. Vis. Exp. (81), e50608, doi:10.3791/50608 (2013).

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