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

Author:
Zhang, Wenli, Li, Caibin, Baguley, Bruce C., Zhou, Fang, Zhou, Weisai, Shaw, John P., Wang, Zhen, Wu, Zimei, Liu, Jianping
Source:
Analytical biochemistry 2016 v.515 pp. 47-54
ISSN:
0003-2697
Subject:
biomimetics, cell suspension culture, centrifugation, collagen, culture media, cytotoxicity, doxorubicin, harvesting, methylcellulose, models, neoplasms, propidium, staining, viability, viscosity
Abstract:
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.
Agid:
5563352