Microfluidic cell culture

Microfluidic cell culture integrates knowledge from biology, biochemistry, engineering, and physics to develop devices and techniques for culturing, maintaining, analyzing, and experimenting with cells at the microscale.[1][2] It merges microfluidics, a set of technologies used for the manipulation of small fluid volumes (μL, nL, pL) within artificially fabricated microsystems, and cell culture, which involves the maintenance and growth of cells in a controlled laboratory environment.[3][4] Microfluidics has been used for cell biology studies as the dimensions of the microfluidic channels are well suited for the physical scale of cells.[2] For example, eukaryotic cells have linear dimensions between 10-100 μm which falls within the range of microfluidic dimensions.[4] A key component of microfluidic cell culture is being able to mimic the cell microenvironment which includes soluble factors that regulate cell structure, function, behavior, and growth.[2] Another important component for the devices is the ability to produce stable gradients that are present in vivo as these gradients play a significant role in understanding chemotactic, durotactic, and haptotactic effects on cells.[2]

Fabrication

Some considerations for microfluidic devices relating to cell culture include:

Fabrication material is crucial as not all polymers are biocompatible, with some materials such as PDMS causing undesirable adsorption or absorption of small molecules.[9][10] Additionally, uncured PDMS oligomers can leach into the cell culture media, which can harm the microenvironment.[9] As an alternative to commonly used PDMS, there have been advances in the use of thermoplastics (e.g., polystyrene) as a replacement material.[11][12]

Spatial organization of cells in microscale devices largely depends on the culture region geometry for cells to perform functions in vivo.[13][14] For example, long, narrow channels may be desired to culture neurons.[13] The perfusion system chosen might also affect the geometry chosen. For example, in a system that incorporates syringe pumps, channels for perfusion inlet, perfusion outlet, waste, and cell loading would need to be added for the cell culture maintenance.[15] Perfusion in microfluidic cell culture is important to enable long culture periods on-chip and cell differentiation.[16]

Other critical aspects for controlling the microenvironment include: cell seeding density, reduction of air bubbles as they can rupture cell membranes, evaporation of media due to an insufficiently humid environment, and cell culture maintenance (i.e. regular, timely media changes).[17][16][18]

Advantages

Some of the major advantages of microfluidic cell culture include reduced sample volumes (especially important when using primary cells, which are often limited) and the flexibility to customize and study multiple microenvironments within the same device.[19] A reduced cell population can also be used in a microscale system (e.g., a few hundred cells) in comparison to macroscale culture systems (which often require 105 – 107 cells); this can make studying certain cell-cell interactions more accessible.[20] These reduced cell numbers make studying non-dividing or slow dividing cells (e.g., stem cells) easier than traditional culture methods (e.g., flasks, petri dishes, or well plates) due to the smaller sample volumes.[20][21] Given the small dimensions in microfluidics, laminar flow can be achieved, allowing manipulations with the culture system to be done easily without affecting other culture chambers.[21] Laminar flow is also useful as is it mimics in vivo fluid dynamics more accurately, often making microscale culture more relevant than traditional culture methods.[22]

Culture Platforms

Two-dimensional culture

Two-dimensional (2D) cell culture is cell culture that takes place on a flat surface, e.g. the bottom of a well-plate, and is known as the conventional method.[23] While these platforms are useful for growing and passaging cells to be used in subsequent experiments, they are not ideal environments to monitor cell responses to stimuli as cells cannot freely move or perform functions as observed in vivo that are dependent on cell-extracellular matrix material interactions.[23]

Three-dimensional culture

Three-dimensional (3D) cell culture is cell culture that takes place in a biologically relevant matrix, usually this involves cells being embedded in a hydrogel containing extracellular molecules (e.g., collagen).[23] By adding an additional dimension, more advanced cell architectures can be achieved, and cell behavior is more representative of in vivo dynamics; cells can engage in enhanced communication with neighboring cells and cell-extracellular matrix interactions can be modeled.[23][24] These simplified 3D cell culture models can be combined in a manner that recapitulates tissue- and organ-level functions in devices known as organ-on-a-chip.[23] In these devices, chambers or collagen layers containing different cell types can interact with one another for multiple days while various channels deliver nutrients to the cells.[23][25] An advantage of these devices is that tissue function can be characterized and observed under controlled conditions (e.g., effect of shear stress on cells, effect of cyclic strain or other forces) to better understand the overall function of the organ.[23][26] While these 3D models often better model organ function on a cellular level compared with 2D models, there are still challenges. Some of the challenges include: imaging of the cells, control of gradients in static models (i.e., without a perfusion system), and difficulty recreating vasculature.[26] Despite these challenges, 3D models are still used as tools for studying and testing drug responses in pharmacological studies.[23]

References

  1. 1 2 Bhatia, Sangeeta N; Ingber, Donald E (August 2014). "Microfluidic organs-on-chips". Nature Biotechnology. 32 (8): 760–772. doi:10.1038/nbt.2989. ISSN 1546-1696. PMID 25093883.
  2. 1 2 3 4 Young, Edmond W. K.; Beebe, David J. (2010-02-24). "Fundamentals of microfluidic cell culture in controlled microenvironments". Chemical Society Reviews. 39 (3): 1036–1048. doi:10.1039/b909900j. ISSN 1460-4744. PMC 2967183. PMID 20179823.
  3. ↑ Mehling, Matthias; Tay, Savaş (2014). "Microfluidic cell culture". Current Opinion in Biotechnology. 25: 95–102. doi:10.1016/j.copbio.2013.10.005. PMID 24484886.
  4. 1 2 Whitesides, George M. (2006-07-26). "The origins and the future of microfluidics". Nature. 442 (7101): 368–373. doi:10.1038/nature05058. PMID 16871203.
  5. ↑ Cho, Brenda S.; Schuster, Timothy G.; Zhu, Xiaoyue; Chang, David; Smith, Gary D.; Takayama, Shuichi (2003-04-01). "Passively Driven Integrated Microfluidic System for Separation of Motile Sperm". Analytical Chemistry. 75 (7): 1671–1675. doi:10.1021/ac020579e. ISSN 0003-2700.
  6. ↑ Zimmermann, Martin; Schmid, Heinz; Hunziker, Patrick; Delamarche, Emmanuel (2006-12-19). "Capillary pumps for autonomous capillary systems". Lab Chip. 7 (1): 119–125. doi:10.1039/b609813d. ISSN 1473-0189. PMID 17180214.
  7. ↑ Walker, Glenn M.; Beebe, David J. (2002-08-21). "A passive pumping method for microfluidic devices". Lab on a Chip. 2 (3): 131–4. CiteSeerX 10.1.1.118.5648. doi:10.1039/b204381e. ISSN 1473-0189. PMID 15100822.
  8. ↑ Kim, Lily; Toh, Yi-Chin; Voldman, Joel; Yu, Hanry (2007-05-30). "A practical guide to microfluidic perfusion culture of adherent mammalian cells". Lab on a Chip. 7 (6): 681–94. doi:10.1039/b704602b. ISSN 1473-0189. PMID 17538709.
  9. 1 2 Regehr, Keil J.; Domenech, Maribella; Koepsel, Justin T.; Carver, Kristopher C.; Ellison-Zelski, Stephanie J.; Murphy, William L.; Schuler, Linda A.; Alarid, Elaine T.; Beebe, David J. (2009-08-07). "Biological implications of polydimethylsiloxane-based microfluidic cell culture". Lab on a Chip. 9 (15): 2132–2139. doi:10.1039/b903043c. ISSN 1473-0189. PMC 2792742. PMID 19606288.
  10. ↑ Halldorsson, Skarphedinn; Lucumi, Edinson; Gómez-Sjöberg, Rafael; Fleming, Ronan M.T. (2015). "Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices". Biosensors and Bioelectronics. 63: 218–231. doi:10.1016/j.bios.2014.07.029. PMID 25105943.
  11. ↑ Berthier, Erwin; Young, Edmond W. K.; Beebe, David (2012-03-07). "Engineers are from PDMS-land, Biologists are from Polystyrenia". Lab on a Chip. 12 (7): 1224–37. doi:10.1039/c2lc20982a. ISSN 1473-0189. PMID 22318426.
  12. ↑ van Midwoud, Paul M.; Janse, Arnout; Merema, Marjolijn T.; Groothuis, Geny M. M.; Verpoorte, Elisabeth (2012-05-01). "Comparison of Biocompatibility and Adsorption Properties of Different Plastics for Advanced Microfluidic Cell and Tissue Culture Models". Analytical Chemistry. 84 (9): 3938–3944. doi:10.1021/ac300771z. ISSN 0003-2700. PMID 22444457.
  13. 1 2 Rhee, Seog Woo; Taylor, Anne M.; Tu, Christina H.; Cribbs, David H.; Cotman, Carl W.; Jeon, Noo Li (2005-12-23). "Patterned cell culture inside microfluidic devices". Lab on a Chip. 5 (1): 102–7. doi:10.1039/b403091e. hdl:10371/7982. ISSN 1473-0189. PMID 15616747.
  14. ↑ Folch, Albert; Toner, Mehmet (1998-01-01). "Cellular Micropatterns on Biocompatible Materials". Biotechnology Progress. 14 (3): 388–392. doi:10.1021/bp980037b. ISSN 1520-6033. PMID 9622519.
  15. ↑ Hung, Paul J.; Lee, Philip J.; Sabounchi, Poorya; Lin, Robert; Lee, Luke P. (2005-01-05). "Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays". Biotechnology and Bioengineering. 89 (1): 1–8. doi:10.1002/bit.20289. ISSN 1097-0290. PMID 15580587.
  16. 1 2 Tourovskaia, Anna; Figueroa-Masot, Xavier; Folch, Albert (2005-12-23). "Differentiation-on-a-chip: A microfluidic platform for long-term cell culture studies". Lab on a Chip. 5 (1): 14–9. doi:10.1039/b405719h. ISSN 1473-0189. PMID 15616734.
  17. ↑ Meyvantsson, Ivar; Beebe, David J. (2008-06-13). "Cell Culture Models in Microfluidic Systems". Annual Review of Analytical Chemistry. 1 (1): 423–449. doi:10.1146/annurev.anchem.1.031207.113042. ISSN 1936-1327. PMID 20636085.
  18. ↑ Yu, Hongmei; Alexander, Caroline M.; Beebe, David J. (2007-05-30). "Understanding microchannel culture: parameters involved in soluble factor signaling". Lab on a Chip. 7 (6): 726–30. doi:10.1039/b618793e. ISSN 1473-0189. PMID 17538714.
  19. ↑ Mehling M, Tay S (February 2014). "Microfluidic cell culture". Current Opinion in Biotechnology. 25: 95–102. doi:10.1016/j.copbio.2013.10.005. PMID 24484886.
  20. 1 2 Halldorsson S, Lucumi E, Gómez-Sjöberg R, Fleming RM (January 2015). "Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices". Biosensors & Bioelectronics. 63: 218–31. doi:10.1016/j.bios.2014.07.029. PMID 25105943.
  21. 1 2 Gómez-Sjöberg R, Leyrat AA, Pirone DM, Chen CS, Quake SR (November 2007). "Versatile, fully automated, microfluidic cell culture system". Analytical Chemistry. 79 (22): 8557–63. doi:10.1021/ac071311w. PMID 17953452.
  22. ↑ Cimetta E, Vunjak-Novakovic G (September 2014). "Microscale technologies for regulating human stem cell differentiation". Experimental Biology and Medicine. 239 (9): 1255–63. doi:10.1177/1535370214530369. PMC 4476254. PMID 24737735.
  23. 1 2 3 4 5 6 7 8 Bhatia SN, Ingber DE (August 2014). "Microfluidic organs-on-chips". Nature Biotechnology. 32 (8): 760–72. doi:10.1038/nbt.2989. PMID 25093883.
  24. ↑ Pampaloni F, Reynaud EG, Stelzer EH (October 2007). "The third dimension bridges the gap between cell culture and live tissue". Nature Reviews. Molecular Cell Biology. 8 (10): 839–45. doi:10.1038/nrm2236. PMID 17684528.
  25. ↑ Huh D, Hamilton GA, Ingber DE (December 2011). "From 3D cell culture to organs-on-chips". Trends in Cell Biology. 21 (12): 745–54. doi:10.1016/j.tcb.2011.09.005. PMC 4386065. PMID 22033488.
  26. 1 2 van Duinen V, Trietsch SJ, Joore J, Vulto P, Hankemeier T (December 2015). "Microfluidic 3D cell culture: from tools to tissue models". Current Opinion in Biotechnology. 35: 118–26. doi:10.1016/j.copbio.2015.05.002. PMID 26094109.
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