Generation of primary hepatocyte microarrays by piezoelectric printing

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• 作者：Alicja  ; Zarowna-Dabrowska^a ;   ; ^b ; Ekaterina O.  ; McKenna^c ; Maaike E.  ; Schutte^d ; Andrew  ; Glidle^a ; Li  ; Chen^a ; Carlos  ; Cuestas-Ayllon^a ; Damian  ; Marshall^d ; Andrew  ; Pitt^c ; Martin D.  ; Dawson^b ; Erdan  ; Gu^b ;   ; ^{erdan.gu@strath.ac.uk} ; Jon M.  ; Cooper^a ; Huabing  ; Yin^a ;   ; ^{huabing.yin@glasgow.ac.uk}
• 关键词：Collagen micro patterning ; Cell micro arrays ; Primary hepatocytes ; Cell viability ; Piezoelectric printing ; Metabolism
• 刊名：Colloids and Surfaces B: Biointerfaces
• 出版年：2012
• 出版时间：1 January 2012
• 年：2012
• 卷：89
• 期：Complete
• 页码：126-132
• 全文大小：841 K

文摘

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.