A biomimetic synthetic feeder layer supports the proliferation and self-renewal of mouse embryonic stem cells

被引:9
作者
Lopez-Fagundo, Cristina [1 ,2 ,6 ]
Livi, Liane L. [1 ]
Ramchal, Talisha [1 ,7 ]
Darling, Eric M. [1 ,2 ,3 ,4 ]
Hoffman-Kim, Diane [1 ,2 ,3 ,5 ]
机构
[1] Brown Univ, Dept Mol Pharmacol Physiol & Biotechnol, 175 Meeting St, Providence, RI 02912 USA
[2] Brown Univ, Ctr Biomed Engn, Providence, RI 02912 USA
[3] Brown Univ, Sch Engn, Providence, RI 02912 USA
[4] Brown Univ, Dept Orthopaed, Providence, RI 02912 USA
[5] Brown Univ, Brown Inst Brain Sci, Providence, RI 02912 USA
[6] Univ Zurich, Ctr Appl Biotechnol & Mol Med, CH-8006 Zurich, Switzerland
[7] Duke Univ, Sch Med, Durham, NC 27710 USA
基金
美国国家科学基金会;
关键词
Embryonic stem cells; Mouse embryonic fibroblast; Feeder layer; Polyacrylamide; DIFFERENTIATION; MATRIX; TOPOGRAPHY; DERIVATION; CULTURE; GROWTH; LINES;
D O I
10.1016/j.actbio.2016.04.047
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
Successful realization of the enormous potential of pluripotent stem cells in regenerative medicine demands the development of well-defined culture conditions. Maintenance of embryonic stem cells (ESCs) typically requires co-culture with feeder layer cells, generally mouse embryonic fibroblasts (MEFs). Concerns about xenogeneic pathogen contamination and immune reaction to feeder cells underlie the need for ensuring the safety and efficacy of future stem cell-based products through the development of a controlled culture environment. To gain insight into the effectiveness of MEF layers, here we have developed a biomimetic synthetic feeder layer (BSFL) that is acellular and replicates the stiffness and topography of MEFs. The mechanical properties of MEFs were measured using atomic force microscopy. The average Young's modulus of the MEF monolayers was replicated using tunable polyacrylamide (PA) gels. BSFLs replicated topographical features of the MEFs, including cellular, subcellular, and cytoskeletal features. On BSFLs, mouse ESCs adhered and formed compact round colonies; similar to on MEF controls but not on Flat PA. ESCs on BSFLs maintained their pluripotency and self-renewal across passages, formed embryoid bodies and differentiated into progenitors of the three germ layers. This acellular biomimetic synthetic feeder layer supports stem cell culture without requiring co-culture of live xenogeneic feeder cells, and provides a versatile, tailorable platform for investigating stem cell growth. Statement of Significance Embryonic stem cells have enormous potential to aid therapeutics, because they can renew themselves and become different cell types. This study addresses a key challenge for ESC use- growing them safely for human patients. ESCs typically grow with a feeder layer of mouse fibroblasts. Since patients have a risk of immune response to feeder layer cells, we have developed a material to mimic the feeder layer and eliminate this risk. We investigated the influence of feeder layer topography and stiffness on mouse ESCs. While the biomimetic synthetic feeder layer contains no live cells, it replicates the stiffness and topography of feeder layer cells. Significantly, ESCs grown on BSFLs retain their abilities to grow and become multiple cell types. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:55 / 64
页数:10
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