Hostname: page-component-848d4c4894-r5zm4 Total loading time: 0 Render date: 2024-06-30T20:12:08.920Z Has data issue: false hasContentIssue false
  • English
  • Français

Generation of cell-laden hydrogel microspheres using 3D printing-enabled microfluidics

Published online by Cambridge University Press:  15 May 2018

Sanika Suvarnapathaki ,
Rafael Ramos ,
Stephen W. Sawyer ,
Shannon McLoughlin ,
Andrew Ramos ,
Sarah Venn  and
Pranav Soman
Sanika Suvarnapathaki
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Rafael Ramos
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Stephen W. Sawyer
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Shannon McLoughlin
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Andrew Ramos
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Sarah Venn
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
Pranav Soman*
Affiliation:
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, New York 13210, USA
*
a)Address all correspondence to this author. e-mail: psoman@syr.edu
Get access

Abstract

3D printing has been shown to be a robust and inexpensive manufacturing tool for a range of applications within biomedical science. Here we report the design and fabrication of a 3D printer-enabled microfluidic device used to generate cell-laden hydrogel microspheres of tunable sizes. An inverse mold was printed using a 3D printer, and replica molding was used to fabricate a PDMS microfluidic device. Intersecting channel geometry was used to generate perfluorodecalin oil-coated gelatin methacrylate (GelMA) microspheres of varying sizes (35–250 μm diameters). Process parameters such as viscosity profile and UV cross-linking times were determined for a range of GelMA concentrations (7–15% w/v). Empirical relationships between flow rates of GelMA and oil phases, microspheres size, and associated swelling properties were determined. For cell experiments, GelMA was mixed with human osteosarcoma Saos-2 cells, to generate cell-laden GelMA microspheres with high long-term viability. This simple, inexpensive method does not require the use of traditional cleanroom facilities and when combined with the appropriate flow setup is robust enough to yield tunable cell-laden hydrogel microspheres for potential tissue engineering applications.

Keywords

biomaterialcellular (material form)polymer
Type
Invited Article
Information
Journal of Materials Research , Volume 33 , Issue 14: Focus Issue: 3D Printing of Biomaterials , 27 July 2018 , pp. 2012 - 2018
Copyright
Copyright © Materials Research Society 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

b)

These authors contributed equally to this work.

References

REFERENCES

Zhao, X., Liu, S., Yildirimer, L., Zhao, H., Ding, R., Wang, H., Cui, W., and Weitz, D.: Injectable stem cell-laden photocrosslinkable microspheres fabricated using microfluidics for rapid generation of osteogenic tissue constructs. Adv. Funct. Mater. 26, 2809 (2016).CrossRefGoogle Scholar
van Duinen, V., Trietsch, S.J., Joore, J., Vulto, P., and Hankemeier, T.: Microfluidic 3D cell culture: From tools to tissue models. Curr. Opin. Biotechnol. 35, 118 (2015).CrossRefGoogle ScholarPubMed
Kang, A., Park, J., Ju, J., Jeong, G.S., and Lee, S-H.: Cell encapsulation via microtechnologies. Biomaterials 35, 2651 (2014).CrossRefGoogle ScholarPubMed
Lu, Y-C., Song, W., An, D., Kim, B.J., Schwartz, R., Wu, M., and Ma, M.: Designing compartmentalized hydrogel microparticles for cell encapsulation and scalable 3D cell culture. J. Mater. Chem. B 3, 353 (2015).CrossRefGoogle Scholar
Yue, K., Trujillo-de Santiago, G., Alvarez, M.M., Tamayol, A., Annabi, N., and Khademhosseini, A.: Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels. Biomaterials 73, 254 (2015).CrossRefGoogle ScholarPubMed
Tan, W.H. and Takeuchi, S.: Monodisperse alginate hydrogel microbeads for cell encapsulation. Adv. Mater. 19, 2696 (2007).CrossRefGoogle Scholar
Dupin, D., Fujii, S., Armes, S.P., Reeve, P., and Baxter, S.M.: Efficient synthesis of sterically stabilized pH-responsive microgels of controllable particle diameter by emulsion polymerization. Langmuir 22, 3381 (2006).CrossRefGoogle ScholarPubMed
Antonietti, M., Bremser, W., Mueschenborn, D., Rosenauer, C., Schupp, B., and Schmidt, M.: Synthesis and size control of polystyrene latices via polymerization in microemulsion. Macromolecules 24, 6636 (1991).CrossRefGoogle Scholar
Kumachev, A., Greener, J., Tumarkin, E., Eiser, E., Zandstra, P.W., and Kumacheva, E.: High-throughput generation of hydrogel microbeads with varying elasticity for cell encapsulation. Biomaterials 32, 1477 (2011).CrossRefGoogle ScholarPubMed
Kim, J.W., Utada, A.S., Fernández-Nieves, A., Hu, Z., and Weitz, D.A.: Fabrication of monodisperse gel shells and functional microgels in microfluidic devices. Angew. Chem. 119, 1851 (2007).CrossRefGoogle Scholar
Workman, V.L., Dunnett, S.B., Kille, P., and Palmer, D.: Microfluidic chip-based synthesis of alginate microspheres for encapsulation of immortalized human cells. Biomicrofluidics 1, 014105 (2007).CrossRefGoogle ScholarPubMed
Li, T., Zhao, L., Liu, W., Xu, J., and Wang, J.: Simple and reusable off-the-shelf microfluidic devices for the versatile generation of droplets. Lab Chip 16, 4718 (2016).CrossRefGoogle ScholarPubMed
Au, A.K., Huynh, W., Horowitz, L.F., and Folch, A.: 3D-printed microfluidics. Angew. Chem., Int. Ed. 55, 3862 (2016).CrossRefGoogle ScholarPubMed
Bhattacharjee, N., Urrios, A., Kang, S., and Folch, A.: The upcoming 3D-printing revolution in microfluidics. Lab Chip 16, 1720 (2016).CrossRefGoogle ScholarPubMed
Kitson, P.J., Rosnes, M.H., Sans, V., Dragone, V., and Cronin, L.: Configurable 3D-printed millifluidic and microfluidic ‘lab on a chip’reactionware devices. Lab Chip 12, 3267 (2012).CrossRefGoogle ScholarPubMed
Waheed, S., Cabot, J.M., Macdonald, N.P., Lewis, T., Guijt, R.M., Paull, B., and Breadmore, M.C.: 3D printed microfluidic devices: Enablers and barriers. Lab Chip 16, 1993 (2016).CrossRefGoogle ScholarPubMed
Albrecht, L.D., Sawyer, S.W., and Soman, P.: Developing 3D scaffolds in the field of tissue engineering to treat complex bone defects. 3D Print. Addit. Manuf. 3, 106 (2016).CrossRefGoogle Scholar
Ogden, K.M., Aslan, C., Ordway, N., Diallo, D., Tillapaugh-Fay, G., and Soman, P.: Factors affecting dimensional accuracy of 3-D printed anatomical structures derived from CT data. J. Digit. Imag. 28, 654 (2015).CrossRefGoogle ScholarPubMed
Yang, L., Shridhar, S.V., Gerwitz, M., and Soman, P.: An in vitro vascular chip using 3D printing-enabled hydrogel casting. Biofabrication 8, 035015 (2016).CrossRefGoogle Scholar
Sawyer, S., Oest, M., Margulies, B., and Soman, P.: Behavior of encapsulated saos-2 cells within gelatin methacrylate hydrogels. J. Tissue Sci. Eng. 7, 2 (2016).CrossRefGoogle Scholar
Chen, Y.X., Yang, S., Yan, J., Hsieh, M-H., Weng, L., Ouderkirk, J.L., Krendel, M., and Soman, P.: A novel suspended hydrogel membrane platform for cell culture. J. Nanotechnol. Eng. Med. 6, 021002 (2015).CrossRefGoogle Scholar
Tamimi, F., Comeau, P., Le Nihouannen, D., Zhang, Y., Bassett, D., Khalili, S., Gbureck, U., Tran, S., Komarova, S., and Barralet, J.: Perfluorodecalin and bone regeneration. Eur. Cell. Mater. 25, 22 (2013).CrossRefGoogle ScholarPubMed
Chokkalingam, V., Weidenhof, B., Krämer, M., Maier, W.F., Herminghaus, S., and Seemann, R.: Optimized droplet-based microfluidics scheme for sol–gel reactions. Lab Chip 10, 1700 (2010).CrossRefGoogle ScholarPubMed
Kumar, D., Gerges, I., Tamplenizza, M., Lenardi, C., Forsyth, N.R., and Liu, Y.: Three-dimensional hypoxic culture of human mesenchymal stem cells encapsulated in a photocurable, biodegradable polymer hydrogel: A potential injectable cellular product for nucleus pulposus regeneration. Acta Biomater. 10, 3463 (2014).CrossRefGoogle Scholar
Chung, C., Mesa, J., Randolph, M.A., Yaremchuk, M., and Burdick, J.A.: Influence of gel properties on neocartilage formation by auricular chondrocytes photoencapsulated in hyaluronic acid networks. J. Biomed. Mater. Res., Part A 77, 518 (2006).CrossRefGoogle ScholarPubMed
Chen, Y.X., Cain, B., and Soman, P.: Gelatin methacrylate-alginate hydrogel with tunable viscoelastic properties. AIMS Mater. Sci. 4, 363 (2017).CrossRefGoogle Scholar
Sawyer, S.W., Dong, P., Venn, S., Ramos, A., Quinn, D., Horton, J.A., and Soman, P.: Conductive gelatin methacrylate-poly(aniline) hydrogel for cell encapsulation. Biomed. Phys. Eng. Express 4, 015005 (2017).CrossRefGoogle Scholar
White, F.M.: Fluid Mechanics (WCB Ed McGraw-Hill, Boston, 1999).Google Scholar
Larson, R.G.: The Structure and Rheology of Complex Fluids (Oxford University Press, New York, 1999).Google Scholar
Agarwal, P., Zhao, S., Bielecki, P., Rao, W., Choi, J.K., Zhao, Y., Yu, J., Zhang, W., and He, X.: One-step microfluidic generation of pre-hatching embryo-like core–shell microcapsules for miniaturized 3D culture of pluripotent stem cells. Lab Chip 13, 4525 (2013).CrossRefGoogle ScholarPubMed