Spheroid Formation from Sorted Cells

Mouse sorted cells at 1 × 10³ were transferred to non-adherent 24 well plates (Corning) with spheroid-forming media^{5 (link), 7 (link), 47 (link)} [1:2 ratio of DMEM F12 (Gibco) and Human Endothelial Medium (Gibco) supplemented with 3.75% Chicken Extract (US Biological), 0.1 mM β-ME (Invitrogen), 1% Non-essential amino acids (Gibco), 1% Pen-strep (Gibco), 1% N2 (Gibco), 2% B27 (Gibco), 20 ng/mL human bFGF (R&D Systems), 20 ng/mL mouse PDGF (Peprotech), 20 ng/mL mouse oncostatin M (R&D Systems), 20 ng/mL mouse IGF-1 (Peprotech), 20 ng/mL mouse EGF (Peprotech)]. After 7 days, the spheroid efficiency was determined.

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Yang M., Arai A., Udagawa N., Hiraga T., Lijuan Z., Ito S., Komori T., Moriishi T., Matsuo K., Shimoda K., Zahalka A.H., Kobayashi Y., Takahashi N, & Mizoguchi T. (2017). Osteogenic Factor Runx2 Marks a Subset of Leptin Receptor-Positive Cells that Sit Atop the Bone Marrow Stromal Cell Hierarchy. Scientific Reports, 7, 4928.

Publication 2017

non-adherent 24 well plates DMEM F12 Human Endothelial Medium Non-essential amino acids Pen-strep human bFGF mouse PDGF mouse oncostatin M mouse EGF

Biological Cells Chicken Endothelial Essential amino acids Human Mouse Mouse oncostatin m Pdgf Strep

Corresponding Organization :

Other organizations : Matsumoto Dental University, Shinshu University, Nagasaki University, Keio University, Albert Einstein College of Medicine

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Variable analysis

independent variables

Mouse sorted cells at 1 × 10^3 concentration

dependent variables

Spheroid efficiency after 7 days

control variables

Non-adherent 24 well plates (Corning)
Spheroid-forming media composition [1:2 ratio of DMEM F12 (Gibco) and Human Endothelial Medium (Gibco) supplemented with 3.75% Chicken Extract (US Biological), 0.1 mM β-ME (Invitrogen), 1% Non-essential amino acids (Gibco), 1% Pen-strep (Gibco), 1% N2 (Gibco), 2% B27 (Gibco), 20 ng/mL human bFGF (R&D Systems), 20 ng/mL mouse PDGF (Peprotech), 20 ng/mL mouse oncostatin M (R&D Systems), 20 ng/mL mouse IGF-1 (Peprotech), 20 ng/mL mouse EGF (Peprotech)]

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