3D Cell Culture Protocols

Key industry publications and methods

There is a diverse range of protocols available in the literature around 3D cell culture and analysis. Discover those highlighted here to learn how researchers use Invitrogen and Gibco products in their methods.

NEWOverview of the cell lines, culture conditions, and analysis techniques that are being used in the transition from 2D to 3D cell models in the 3D Cell Culture & Analysis Handbook.

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NEWHave fun with mazes, lab awards, a word search, and more in the 3D Cell Culture & Analysis Activity Booklet.

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FREE 3D eLearning Courses

We are now offering an ever-expanding set of free e-Learns to pass on the expertise and assist with advancing your research. Follow the links below and start your free learning adventure in the basics of 3D and then progress to our neural organoid training course.

Getting started in 3D cell culture
Learn more at: thermofisher.com/3dbasics-elearn

Neural organoid generation from PSCs
Learn more at: thermofisher.com/neuralorganoid-elearn

Brain

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	FGF2	DMEM/F12, Neurobasal	Lancaster, M.A., et al. Nature. 2013
Pluripotent stem cells	FGF2, EGF, BDNF, NT-3	DMEM/F12, Neurobasal	Pasca, A.M. et al. Nature Methods. 2015
Pluripotent stem cells	DKK1	DMEM/F12	Phillips, A.W., et al. J. Vis. Exp. 2017
Pluripotent stem cells	DKK1, BMPR1a-Fc	DMEM/F12, Neurobasal	Mariani, J., et al. PNAS. 2012
Pluripotent stem cells	N/A	DMEM/F12, Neurobasal	Iefremova, V., et al., Cell Reports. 2017
Pluripotent stem cells	BDNF, GDNF, NT-3, Laminin, bFGF	DMEM/F12, Essential 8, Essential 6, Neurobasal	Zhou, T., et al., Cell Stem Cell. 2017

Liver

Cell type	Relevant growth factors	Relevant media	Key publication
Adult tissue stem/progenitor cells	FGF10, HGF, EGF, Noggin	Advanced DMEM/F12	Huch, M., et al. Nature. 2013
Adult tissue stem/progenitor cells	EGF, FGF10, HGF, Noggin, BMP7, FGF19	Advanced DMEM/F12	Huch, M., et al. Cell 2015
Pluripotent stem cells	BMP4, VEGF, Activin, PDGFBB, FGF2	RPMI1640, DMEM/F12, StemPro-34 SFM	Takebe, T., et al. Cell Reports. 2017
Pluripotent stem cells	FGF2, BMP4, Activin, HGF, EGF	RPMI1640	Takebe, T., et al. Nature. 2013
Primary cells	N/A	HepaRG media	Proctor, W.R., et al. Arch. Toxicol. 2017

Intestine

Cell type	Relevant growth factors	Relevant media	Key publication
Adult tissue stem/progenitor cells	Noggin, EGF, FGF10	DMEM, Advanced DMEM/F12	Sato, T, et al., Gastroenterology. 2011
Adult tissue stem/progenitor cells	EGF, Noggin	Advanced DMEM/F12	Sato, T., et al., Nature, 2009
Pluripotent stem cells	Activin, FGF4, BMP2, EGF, Noggin, SHH	N/A	Munera, J.O., et al., Cell Stem Cell. 2017
Pluripotent stem cells	Activin, FGF4, Noggin, EGF	DMEM/F12, RPMI1640, Advanced DMEM/F12	Spence, J.R., et al., Nature. 2011
Pluripotent stem cells	IGF1, FGF2	Knockout DMEM	Uchida, H., et al., JCI Insight. 2016
Pluripotent stem cells	Activin, FGF4, EGF, Noggin	RPMI1640, Advanced DMEM/F12	Watson, C.L., et al., Nature Medicine. 2014

Kidney

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	FGF2, Noggin, Activin, FGF9	Advanced RPMI1640	Morizane, R., et al., Nature Biotechnology. 2015
Pluripotent stem cells	FGF2, Noggin, Activin, FGF9	Advanced RPMI1640	Freedman, B.S., et al., Nature Communications. 2015
Pluripotent stem cells	FGF2, BMP4, Activin, BMP7, FGF9	DMEM/F12	Takasato, M., et al., Nautre. 2014
Fetal tissue-derived cells	BMP7, FGF2, LIF	DMEM/F12	Li, Z., et al., Cell Stem Cell. 2016

Lung

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	Activin, FGF4, FGF2, Noggin, SHH	RPMI1640, Advanced DMEM/F12	Dye, BR., et al., Elife. 2015
Pluripotent stem cells	FGF2	Advanced DMEM/F12	Wilkinson, DC., et al., Stem Cells Translational Medicine. 2016

Brain

Brain

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	FGF2	DMEM/F12, Neurobasal	Lancaster, M.A., et al. Nature. 2013
Pluripotent stem cells	FGF2, EGF, BDNF, NT-3	DMEM/F12, Neurobasal	Pasca, A.M. et al. Nature Methods. 2015
Pluripotent stem cells	DKK1	DMEM/F12	Phillips, A.W., et al. J. Vis. Exp. 2017
Pluripotent stem cells	DKK1, BMPR1a-Fc	DMEM/F12, Neurobasal	Mariani, J., et al. PNAS. 2012
Pluripotent stem cells	N/A	DMEM/F12, Neurobasal	Iefremova, V., et al., Cell Reports. 2017
Pluripotent stem cells	BDNF, GDNF, NT-3, Laminin, bFGF	DMEM/F12, Essential 8, Essential 6, Neurobasal	Zhou, T., et al., Cell Stem Cell. 2017

Liver

Liver

Cell type	Relevant growth factors	Relevant media	Key publication
Adult tissue stem/progenitor cells	FGF10, HGF, EGF, Noggin	Advanced DMEM/F12	Huch, M., et al. Nature. 2013
Adult tissue stem/progenitor cells	EGF, FGF10, HGF, Noggin, BMP7, FGF19	Advanced DMEM/F12	Huch, M., et al. Cell 2015
Pluripotent stem cells	BMP4, VEGF, Activin, PDGFBB, FGF2	RPMI1640, DMEM/F12, StemPro-34 SFM	Takebe, T., et al. Cell Reports. 2017
Pluripotent stem cells	FGF2, BMP4, Activin, HGF, EGF	RPMI1640	Takebe, T., et al. Nature. 2013
Primary cells	N/A	HepaRG media	Proctor, W.R., et al. Arch. Toxicol. 2017

Intestine

Intestine

Cell type	Relevant growth factors	Relevant media	Key publication
Adult tissue stem/progenitor cells	Noggin, EGF, FGF10	DMEM, Advanced DMEM/F12	Sato, T, et al., Gastroenterology. 2011
Adult tissue stem/progenitor cells	EGF, Noggin	Advanced DMEM/F12	Sato, T., et al., Nature, 2009
Pluripotent stem cells	Activin, FGF4, BMP2, EGF, Noggin, SHH	N/A	Munera, J.O., et al., Cell Stem Cell. 2017
Pluripotent stem cells	Activin, FGF4, Noggin, EGF	DMEM/F12, RPMI1640, Advanced DMEM/F12	Spence, J.R., et al., Nature. 2011
Pluripotent stem cells	IGF1, FGF2	Knockout DMEM	Uchida, H., et al., JCI Insight. 2016
Pluripotent stem cells	Activin, FGF4, EGF, Noggin	RPMI1640, Advanced DMEM/F12	Watson, C.L., et al., Nature Medicine. 2014

Kidney

Kidney

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	FGF2, Noggin, Activin, FGF9	Advanced RPMI1640	Morizane, R., et al., Nature Biotechnology. 2015
Pluripotent stem cells	FGF2, Noggin, Activin, FGF9	Advanced RPMI1640	Freedman, B.S., et al., Nature Communications. 2015
Pluripotent stem cells	FGF2, BMP4, Activin, BMP7, FGF9	DMEM/F12	Takasato, M., et al., Nautre. 2014
Fetal tissue-derived cells	BMP7, FGF2, LIF	DMEM/F12	Li, Z., et al., Cell Stem Cell. 2016

Lung

Lung

Cell type	Relevant growth factors	Relevant media	Key publication
Pluripotent stem cells	Activin, FGF4, FGF2, Noggin, SHH	RPMI1640, Advanced DMEM/F12	Dye, BR., et al., Elife. 2015
Pluripotent stem cells	FGF2	Advanced DMEM/F12	Wilkinson, DC., et al., Stem Cells Translational Medicine. 2016

Protocols for spheroid generation

To help guide you in your 3D cell culture, our R&D teams have developed suggested protocols for the generation of spheroids from the most commonly used cancer cell lines using Nunclon Sphera cell culture plastics and Gibco media.