Inducing Non-Myeloid Differentiation in EBs

In order to induce non-myeloid differentiation at day 4, the formed EBs were transferred to T75 flasks in factory medium (FM) X-VIVO 15™ (Lonza) supplemented with GlutaMax 1% (Gibco), b-mercaptoethanol (Gibco), M-CSF 100 ng/mL (PeproTech) and IL-3 25 ng/mL (PeproTech). After day 14, medium was refreshed every 4 days. At day 22 onwards, iMϕ precursors were isolated and plated on 6-well plates in order to differentiate into iMφs. The differentiation medium (DM) used for the terminal differentiation is X-VIVO 15™ (Lonza) supplemented with GlutaMax 1% (Gibco), b-mercaptoethanol (Gibco), and M-CSF 100 ng/mL (PeproTech) for 7 days^{30 (link)}.

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Poulis N., Martin M., Hoerstrup S.P., Emmert M.Y, & Fioretta E.S. (2024). Development of an iPSC-derived tissue-resident macrophage-based platform for the in vitro immunocompatibility assessment of human tissue engineered matrices. Scientific Reports, 14, 12171.

Publication 2024

X-VIVO 15™ GlutaMax b-mercaptoethanol M-CSF IL-3

Corresponding Organization :

Other organizations : Swiss Institute for Regenerative Medicine, University of Zurich

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Variable analysis

independent variables

Day of transfer to T75 flasks in factory medium (FM) X-VIVO 15™ (Lonza) supplemented with GlutaMax 1% (Gibco), b-mercaptoethanol (Gibco), M-CSF 100 ng/mL (PeproTech) and IL-3 25 ng/mL (PeproTech)
Duration of culture in the differentiation medium (DM) X-VIVO 15™ (Lonza) supplemented with GlutaMax 1% (Gibco), b-mercaptoethanol (Gibco), and M-CSF 100 ng/mL (PeproTech)

dependent variables

Non-myeloid differentiation of the formed EBs
Isolation and plating of iMϕ precursors
Differentiation of iMφs

control variables

Type of culture medium (FM and DM)
Concentration of GlutaMax (1%), b-mercaptoethanol, M-CSF (100 ng/mL), and IL-3 (25 ng/mL)

positive controls

Not specified

negative controls

Not specified

Annotations

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