Optimization of the formation of embedded multicellular spheroids of MCF-7 cells: How to reliably produce a biomimetic 3D model

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• 作者：Wenli Zhang^a ; ¹ ; Caibin Li^a ; ¹ ; Bruce C. Baguley^c ; Fang Zhou^d ; Weisai Zhou^a ; John P. Shaw^b ; Zhen Wang^a ; Zimei Wu^b ; ^{z.wu@auckland.ac.nz" class="auth_mail" title="E-mail the corresponding author} ; Jianping Liu^a ; ^{liujianpingljp@hotmail.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：MCF-7 cells ; Multicellular spheroids ; Hanging drop ; Liquid overlay ; Collagen gels
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 December 2016
• 年：2016
• 卷：515
• 期：Complete
• 页码：47-54
• 全文大小：1000 K

文摘

To obtain a multicellular MCF-7 spheroid model to mimic the three-dimensional (3D) of tumors, the microwell liquid overlay (A) and hanging-drop/agar (B) methods were first compared for their technical parameters. Then a method for embedding spheroids within collagen was optimized. For method A, centrifugation assisted cells form irregular aggregates but not spheroids. For method B, an extended sedimentation period of over 24 h for cell suspensions and increased viscosity of the culture medium using methylcellulose were necessary to harvest a dense and regular cell spheroid. When the number was less than 5000 cells/drop, embedded spheroids showed no tight cores and higher viability than the unembedded. However, above 5000 cells/drop, cellular viability of embedded spheroids was not significantly different from unembedded spheroids and cells invading through the collagen were in a sun-burst pattern with tight cores. Propidium Iodide staining indicated that spheroids had necrotic cores. The doxorubicin cytotoxicity demonstrated that spheroids were less susceptible to DOX than their monolayer cells. A reliable and reproducible method for embedding spheroids using the hanging-drop/agarose method within collagen is described herein. The cell culture model can be used to guide experimental manipulation of 3D cell cultures and to evaluate anticancer drug efficacy.