Generation of primary hepatocyte microarrays by piezoelectric printing

Alicja Zarowna-Dabrowska, Ekaterina O. McKenna, Maaike E. Schütte, Andrew Glidle, Li Chen, Carlos Cuestas-Ayllon, Damian Marshall, Andrew Pitt, Martin D Dawson, Erdan Gu, Jon M. Cooper, Huabing Yin

Research output: Contribution to journalArticle

Abstract

We demonstrate a single-step method for the generation of collagen and poly-l-Lysine (PLL) micropatterns on a poly(ethylene glycol) (PEG) functionalized glass surface for cell based assays. The method involves establishing a reliable silanization method to create an effective non-adhesive PEG layer on glass that inhibits cell attachment, followed by the spotting of collagen or PLL solutions using non-contact piezoelectric printing. We show for the first time that the spotted protein micropatterns remain stable on the PEG surface even after extensive washing, thus significantly simplifying protein pattern formation. We found that adherence and spreading of NIH-3T3 fibroblasts was confined to PLL and collagen areas of the micropatterns. In contrast, primary rat hepatocytes adhered and spread only on collagen micropatterns, where they formed uniform, well defined functionally active cell arrays. The differing affinity of hepatocytes and NIH-3T3 fibroblasts for collagen and PLL patterns was used to develop a simple technique for creating a co-culture of the two cell types. This has the potential to form structured arrays that mimic the in vivo hepatic environment and is easily integrated within a miniaturized analytical platform for developing high throughput toxicity analysis in vitro.
LanguageEnglish
Pages126-132
Number of pages7
JournalColloids and Surface B: Biointerfaces
Volume89
Issue number1
Early online date17 Sep 2011
DOIs
Publication statusPublished - 1 Jan 2012

Fingerprint

Printing
collagens
Microarrays
Collagen
printing
lysine
Hepatocytes
Polyethylene glycols
Lysine
fibroblasts
Fibroblasts
cells
Glass
proteins
Proteins
Metrorrhagia
Ethylene Glycol
glass
washing
Coculture Techniques

Keywords

  • animals
  • hepatocytes
  • mice
  • NIH 3T3 cells

Cite this

Zarowna-Dabrowska, A., McKenna, E. O., Schütte, M. E., Glidle, A., Chen, L., Cuestas-Ayllon, C., ... Yin, H. (2012). Generation of primary hepatocyte microarrays by piezoelectric printing. Colloids and Surface B: Biointerfaces, 89(1), 126-132. https://doi.org/10.1016/j.colsurfb.2011.09.016
Zarowna-Dabrowska, Alicja ; McKenna, Ekaterina O. ; Schütte, Maaike E. ; Glidle, Andrew ; Chen, Li ; Cuestas-Ayllon, Carlos ; Marshall, Damian ; Pitt, Andrew ; Dawson, Martin D ; Gu, Erdan ; Cooper, Jon M. ; Yin, Huabing. / Generation of primary hepatocyte microarrays by piezoelectric printing. In: Colloids and Surface B: Biointerfaces. 2012 ; Vol. 89, No. 1. pp. 126-132.
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Zarowna-Dabrowska, A, McKenna, EO, Schütte, ME, Glidle, A, Chen, L, Cuestas-Ayllon, C, Marshall, D, Pitt, A, Dawson, MD, Gu, E, Cooper, JM & Yin, H 2012, 'Generation of primary hepatocyte microarrays by piezoelectric printing' Colloids and Surface B: Biointerfaces, vol. 89, no. 1, pp. 126-132. https://doi.org/10.1016/j.colsurfb.2011.09.016

Generation of primary hepatocyte microarrays by piezoelectric printing. / Zarowna-Dabrowska, Alicja; McKenna, Ekaterina O.; Schütte, Maaike E.; Glidle, Andrew; Chen, Li; Cuestas-Ayllon, Carlos; Marshall, Damian; Pitt, Andrew; Dawson, Martin D; Gu, Erdan; Cooper, Jon M.; Yin, Huabing.

In: Colloids and Surface B: Biointerfaces, Vol. 89, No. 1, 01.01.2012, p. 126-132.

Research output: Contribution to journalArticle

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AU - McKenna, Ekaterina O.

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AU - Chen, Li

AU - Cuestas-Ayllon, Carlos

AU - Marshall, Damian

AU - Pitt, Andrew

AU - Dawson, Martin D

AU - Gu, Erdan

AU - Cooper, Jon M.

AU - Yin, Huabing

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N2 - We demonstrate a single-step method for the generation of collagen and poly-l-Lysine (PLL) micropatterns on a poly(ethylene glycol) (PEG) functionalized glass surface for cell based assays. The method involves establishing a reliable silanization method to create an effective non-adhesive PEG layer on glass that inhibits cell attachment, followed by the spotting of collagen or PLL solutions using non-contact piezoelectric printing. We show for the first time that the spotted protein micropatterns remain stable on the PEG surface even after extensive washing, thus significantly simplifying protein pattern formation. We found that adherence and spreading of NIH-3T3 fibroblasts was confined to PLL and collagen areas of the micropatterns. In contrast, primary rat hepatocytes adhered and spread only on collagen micropatterns, where they formed uniform, well defined functionally active cell arrays. The differing affinity of hepatocytes and NIH-3T3 fibroblasts for collagen and PLL patterns was used to develop a simple technique for creating a co-culture of the two cell types. This has the potential to form structured arrays that mimic the in vivo hepatic environment and is easily integrated within a miniaturized analytical platform for developing high throughput toxicity analysis in vitro.

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Zarowna-Dabrowska A, McKenna EO, Schütte ME, Glidle A, Chen L, Cuestas-Ayllon C et al. Generation of primary hepatocyte microarrays by piezoelectric printing. Colloids and Surface B: Biointerfaces. 2012 Jan 1;89(1):126-132. https://doi.org/10.1016/j.colsurfb.2011.09.016