Constrained spheroids for prolonged hepatocyte culture

详细信息    查看全文

• 作者：Wen Hao Tong^a ; ^b ; ^c ; Yu Fang^a ; ^b ; ^c ; Jie Yan^a ; ^c ; ^g ; Xin Hong^c ; Nisha Hari Singh^a ; Shu Rui Wang^d ; Bramasta Nugraha^a ; ^b ; ^e ; ^f ; Lei Xia^c ; Eliza Li Shan Fong^c ; Ciprian Iliescu^a ; ^{ciliescu@ibn.a-star.edu.sg" class="auth_mail" title="E-mail the corresponding author} ; Hanry Yu^a ; ^b ; ^c ; ^g ; ^h ; ⁱ ; ^{hanry_yu@nuhs.edu.sg" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Hepatocytes ; Spheroids ; Liver ; Perfusion ; Drug testing
• 刊名：Biomaterials
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：80
• 期：Complete
• 页码：106-120
• 全文大小：4428 K

文摘

Liver-specific functions in primary hepatocytes can be maintained over extended duration in vitro using spheroid culture. However, the undesired loss of cells over time is still a major unaddressed problem, which consequently generates large variations in downstream assays such as drug screening. In static culture, the turbulence generated by medium change can cause spheroids to detach from the culture substrate. Under perfusion, the momentum generated by Stokes force similarly results in spheroid detachment. To overcome this problem, we developed a Constrained Spheroids (CS) culture system that immobilizes spheroids between a glass coverslip and an ultra-thin porous Parylene C membrane, both surface-modified with poly(ethylene glycol) and galactose ligands for optimum spheroid formation and maintenance. In this configuration, cell loss was minimized even when perfusion was introduced. When compared to the standard collagen sandwich model, hepatocytes cultured as CS under perfusion exhibited significantly enhanced hepatocyte functions such as urea secretion, and CYP1A1 and CYP3A2 metabolic activity. We propose the use of the CS culture as an improved culture platform to current hepatocyte spheroid-based culture systems.