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Soft fibrin gels promote selection and growth of tumorigenic cells

Abstract

The identification of stem-cell-like cancer cells through conventional methods that depend on stem cell markers is often unreliable. We developed a mechanical method for selecting tumorigenic cells by culturing single cancer cells in fibrin matrices of ~100 Pa in stiffness. When cultured within these gels, primary human cancer cells or single cancer cells from mouse or human cancer cell lines grew within a few days into individual round colonies that resembled embryonic stem cell colonies. Subcutaneous or intravenous injection of 10 or 100 fibrin-cultured cells in syngeneic or severe combined immunodeficiency mice led to the formation of solid tumours at the site of injection or at the distant lung organ much more efficiently than control cancer cells selected using conventional surface marker methods or cultured on conventional rigid dishes or on soft gels. Remarkably, as few as ten such cells were able to survive and form tumours in the lungs of wild-type non-syngeneic mice.

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Figure 1: Multicellular tumour spheroid formation in soft 3D fibrin gels.
Figure 2: Tumour metastasis of 3D-cultured B16-F1 cells in lung tissue of BALB/c mice.
Figure 3: Upregulation of stem-cell-associated genes in B16-F1 spheroid cells cultured in 3D fibrin gels.
Figure 4: 3D-fibrin-gel-cultured B16-F1 cells do not stiffen but elevate tractions with substrate rigidity.

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Acknowledgements

We would like to thank J. Winer for help with fibrin-gel protocols. This work was supported by funds from Huazhong University of Science and Technology (HUST), the National Basic Research Program of China (2012CB932500) (B.H.), the China Natural National Science Foundation (30911120482) (B.H.), the Fundamental Research Funds for the Central Universities (HUST-2011TS027) (B.H.), and USA NIH grant GM072744 (N.W.).

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N.W. and B.H. conceived the project; B.H., N.W., J.L. and Y.T. designed the experiments. B.H., J.L., Y.T., H.Z., Y.Z., P.X., J.C., Y-C.P. and K.T. carried out the experiments and analysed the data. N.W., B.H., J.L. and Y.T. wrote the manuscript.

Corresponding authors

Correspondence to Ning Wang or Bo Huang.

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The authors declare no competing financial interests.

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Liu, J., Tan, Y., Zhang, H. et al. Soft fibrin gels promote selection and growth of tumorigenic cells. Nature Mater 11, 734–741 (2012). https://doi.org/10.1038/nmat3361

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