In a study published last October in the journal Stem Cell Research, Chandrasekaran et al. compared the induction of neural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs) obtained by 2D and 3D culture methods. Neural differentiation of five hiPSC lines was analyzed at the protein level by immunocytochemistry and flow cytometry to reveal the expression of various markers : NPC, neuronal, cortical layer and glial markers.

NPCs derived from human PSCs are often induced using 3D aggregates, because these systems are considered as efficient differentiation tools to induce PSCs into the neural lineage. However, according to the authors, no publication had directly compared 3D neural induction (sphere) with 2D neural induction (monolayer) in terms of cell numbers, gene expression profiles, proliferation rates, differentiation potential, cell fate and functionality.

They found no significant difference between the two induction methods – or the different genetic background cell lines – regarding early differentiation events (neuroepithelial commitment and formation of neural rosettes). However, a higher number of NPCs positive for NESTIN and PAX6 was generated using 3D neural induction, compared to the 2D method. Interestingly, the analysis of neurite length revealed that the 3D-induced neurons exhibited longer neurites than 2D-induced cells.

This study, among others, shows that 3D culture methods might help to improve the differentiation of human cells in vitro, leading to more physiological phenotypes !

It’s always funny to see that the “old” maximum intensity projection is so used in picture reconstruction. Even if we all know artefacts created by this method, we often prefer a parameter free tool … In this article, a new way to do the 3D to 2D reduction is presented, the smooth manifold extraction, and it is compared to others one.

Three-dimensional fluorescence microscopy followed by image processing is routinely used to study biological objects at various scales such as cells and tissue. However, maximum intensity projection, the most broadly used rendering tool, extracts a discontinuous layer of voxels, obliviously creating important artifacts and possibly misleading interpretation. Here we propose smooth manifold extraction, an algorithm that produces a continuous focused 2D extraction from a 3D volume, hence preserving local spat

Source: Smooth 2D manifold extraction from 3D image stack | Nature Communications

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