Skip to main content

Advertisement

SpringerLink
Log in

Use of a gradient-generating microfluidic device to rapidly determine a suitable glucose concentration for cell viability test

  • Original Research
  • Published:
Microfluidics and Nanofluidics Aims and scope Submit manuscript

Abstract

We successfully determined a suitable glucose concentration for endothelial cells (ECs) using a gradient-generating microfluidic chip and a micro-stamper that were fabricated using micro-electro-mechanical systems (MEMS) technology. Our strategy was to generate a stable concentration gradient in the observation area based on a microfluidic network and micro-mixers, which produced a concentration gradient under various flow rates. The areas for cell adhesion were delineated on a glass slide with a micro-stamper using the micro-contact printing (μCP) method. We also discuss which glucose concentration gradients are suitable for cell viability test (i.e., 0–0.2%, 0.05–0.15%, and 0.06–0.17%). After examining various concentration gradients, the suitable glucose concentration for EC’s viability test was determined to range from 0.077% (4.2 mM) to 0.147% (8.16 mM). Higher or lower concentrations caused the ECs to atrophy or die. In this study, we describe a gradient-generating microfluidic chip that can be used to produce various drug concentrations for multi-concentration tests.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Baumgartner-Parzer SM, Wagner L, Pettermann M, Grillari J, Gessl A, Waldhausl W (1995) High-glucose-triggered apoptosis in cultured endothelial cells. Am Diabetes Assoc 44:1323–1327

    Google Scholar 

  • Boyden SV (1962) Rapid quantitation of neutrophil chemotaxis: use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J Immunol Methods 115:453–466

    Google Scholar 

  • Branch DW, Corey JM, Weyhenmeyer JA, Brewer GJ, Wheeler BC (1998) Microstamp patterns of biomolecules for high-resolution neuronal networks. Med Biol Eng Comput 36:135–141

    Article  Google Scholar 

  • Chang JC, Brewer GJ, Wheeler BC (2003) A modified microstamping technique enhances polylysine transfer and neuronal cell patterning. Biomaterials 24:2863–2870

    Article  Google Scholar 

  • Chung BG, Park JW, Hu JS, Huang C, Monuki ES, Jeon NL (2007) A hybrid microfluidic-vacuum device for direct interfacing with conventional cell culture methods. BMC Biotechnol 7:60–67

    Article  Google Scholar 

  • Das T, Mallick SK, Paul D, Bhutia SK, Bhattacharyya TK, Maiti TK (2007) Microcontact printing of concanavalin a and its effect on mammalian cell morphology. J Colloid Interface Sci 314:71–79

    Article  Google Scholar 

  • DCCT Research Group (1993) The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. N Engl J Med 329:977–986

    Article  Google Scholar 

  • Dertinger KWS, Chiu DT, Jeon NL, Whitesides GM (2001) Generation of gradients having complex shapes using microfluidic networks. Anal Chem 73:1240–1246

    Article  Google Scholar 

  • Dewey CF, Bussolari SR, Gimbrone MA, Davies PF (1981) The dynamic response of vascular endothelial cells to fluid shear stress. J Biomech Eng 103:177–185

    Article  Google Scholar 

  • Folch A, Toner M (2000) Microengineering of cellular interactions. Annu Rev Biomed Eng 2:227–256

    Article  Google Scholar 

  • Folch A, Jo BH, Hurtado O, Beebe DJ, Toner M (2000) Microfabricated elastomeric stencils for micropatterning cell cultures. J Biomed Mater Res 52:346–353

    Article  Google Scholar 

  • Jeon NL, Dertinger SKW, Chiu DT, Choi IS, Stroock AD, Whitesides GM (2000) Generation of solution and surface gradients using microfluidic systems. Langmuir 16:8311–8316

    Article  Google Scholar 

  • Katanosaka Y, Bao JH, Komatsu T, Suemori T, Yamada A, Mohri S, Naruse K (2008) Analysis of cyclic-stretching responses using cell-adhesion-patterned cell. J Biotechnol 133:82–89

    Google Scholar 

  • Kim YD, Park CB, Clark DS (2001) Stable sol-gel microstructured and microfluidic networks for protein patterning. Biotechnol Bioeng 73:331–337

    Article  Google Scholar 

  • Kim L, Vahey MD, Lee HY, Voldman J (2006) Microfluidic arrays for logarithmically perfused embryonic stem cell culture. Lab Chip 6:394–406

    Article  Google Scholar 

  • Li Y, Yuan B, Ji H, Han D, Chen S, Tian F, Jiang X (2007) A method for patterning multiple types of cells by using electrochemical desorption of self-assembled monolayers within microfluidic channels. Angew Chem Int Ed 46:1094–1096

    Article  Google Scholar 

  • McGinn S, Poronnik P, King M, Gallery EDM, Pollock CA (2003) High glucose and endothelial cell growth: novel effects independent of autocrine TGF-beta 1 and hyperosmolarity. Am J Physiol Cell Physiol 284:1374–1386

    Google Scholar 

  • Millioni R, Puricelli L, Iori E, Arrigoni G, Tessari P (2010) The effects of rosiglitazone and high glucose on protein expression in endothelial cells. J Proteome Res 9:578–584

    Article  Google Scholar 

  • Nelson RD, Quie PG, Simmons RL (1975) Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leukocytes and monocytes. J Immunol 115:1650–1656

    Google Scholar 

  • Park TH, Shuler ML (2003) Integration of cell culture and microfabrication technology. Biotechnol Prog 19:243–253

    Article  Google Scholar 

  • Rossetti L, Giaccari A, DeFronzo RA (1990) Glucose toxicity. Am Diabetes Assoc 13:610–630

    Google Scholar 

  • Somersalo K, Salob OP, Bjorkstenb F, Mustakallioc KK (1990) A simplified Boyden chamber assay for neutrophil chemotaxis based on quantitation of myeloperoxidase. Anal Biochem 185:238–242

    Article  Google Scholar 

  • Song H, Poo MM (1999) Signal transduction underlying growth cone guidance by diffusible factors. Curr Opin Neurobiol 9:355–363

    Article  Google Scholar 

  • Takayama S, McDonald JC, Ostuni E, Liang MN, Kenis PJA, Ismagilov RF, Whitesides GM (1999) Patterning cells and their environments using multiple laminar fluid flows in capillary networks. Proc Natl Acad Sci USA 96:5545–5548

    Article  Google Scholar 

  • Takayama S, Ostuni E, Qian XP, McDonald JC, Jiang XY, LeDuc P, Wu MH, Ingber DE, Whitesides GM (2001a) Topographical micropatterning of poly(dimethylsiloxane) using laminar flows of liquids in capillaries. Adv Mater 13:570–574

    Article  Google Scholar 

  • Takayama S, Ostuni E, LeDuc P, Naruse K, Ingber DE, Whitesides GM (2001b) Laminar flows-subcellular positioning of small molecules. Nature 411:1016

    Article  Google Scholar 

  • Wang S, Yue F, Zhang L, Wang J, Wang Y, Jiang L, Lin B, Wang Q (2009) Simultaneous quantification of active components in the herbs and products of Si-Wu-Tang by high performance liquid chromatography-mass spectrometry. J Pharm Biomed Anal 49:806–810

    Article  Google Scholar 

  • Wechezak AR, Viggers RF, Sauvage LR (1985) Fibronectin and F-actin redistribution in cultured endothelial cells exposed to shear stress. Lab Invest 53:639–647

    Google Scholar 

  • Wyart C, Ybert C, Bourdieu L, Herr C, Prinz C, Chatenay D (2002) Constrained synaptic connectivity in functional mammalian neuronal networks grown on patterned surfaces. J Neurosci Methods 117:123–131

    Article  Google Scholar 

  • Yki-Järvinen H, Helve E, Koivisto VA (1987) Hyperglycemia decreases glucose uptake in type I diabetes. Am Diabetes Assoc 36:892–896

    Google Scholar 

Download references

Acknowledgments

The authors would like to thank the Center for Micro/Nano Technology, National Cheng Kung University, Tainan, Taiwan, R.O.C., for access to equipment and technical support. Funding from the Ministry of Education and the National Science Council of Taiwan, R.O.C. under Grants NSC 97-2221-E-006-222-MY3 and NSC 99-2221-E-006-203-MY3 are gratefully acknowledged.

Author information

Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, 1 University Road, Tainan, 701, Taiwan, ROC

    Chia-Hsien Yeh, Chien-Hsien Chen & Yu-Cheng Lin

  2. Center for Micro/Nano Technology, National Cheng Kung University, Tainan, Taiwan, ROC

    Yu-Cheng Lin

Authors
  1. Chia-Hsien Yeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chien-Hsien Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yu-Cheng Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yu-Cheng Lin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yeh, CH., Chen, CH. & Lin, YC. Use of a gradient-generating microfluidic device to rapidly determine a suitable glucose concentration for cell viability test. Microfluid Nanofluid 10, 1011–1018 (2011). https://doi.org/10.1007/s10404-010-0730-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10404-010-0730-0

Keywords

Search

Navigation