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Université de Strasbourg, INSERM, EFS Grand Est

6 ARTICLES PUBLISHED IN JoVE

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Biology

In Situ Exploration of Murine Megakaryopoiesis using Transmission Electron Microscopy
Cyril Scandola 1, François Lanza 1, Christian Gachet 1, Anita Eckly 1
1Université de Strasbourg, INSERM, EFS Grand Est, BPPS UMR-S 1255, FMTS

Here, we present a protocol to analyze ultrastructure of the megakaryocytes in situ using transmission electron microscopy (TEM). Murine bone marrows are collected, fixed, embedded in epoxy resin and cut in ultrathin sections. After contrast staining, the bone marrow is observed under a TEM microscope at 120 kV.

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Biology

Isolation of Mouse Megakaryocyte Progenitors
Quentin Kimmerlin 1, Manuela Tavian 2, Christian Gachet 1, François Lanza 1, Nathalie Brouard 1
1Université de Strasbourg, INSERM UMR S1255, EFS Grand-EST, 2UMR-S1113 –IRFAC, Université de Strasbourg

This method describes the purification by flow cytometry of MEP and MKp from mice femurs, tibias, and pelvic bones.

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Biology

Leukodepletion Filters-Derived CD34+ Cells As a Cell Source to Study Megakaryocyte Differentiation and Platelet Formation
Anaïs Pongerard 1, Lea Mallo 1, Christian Gachet 1, Henri de La Salle 1, François Lanza 1, Catherine Strassel 1
1BPPS UMR-S 1255, FMTS, Université de Strasbourg, INSERM, EFS Grand Est

This protocol describes in detail all the steps involved in obtaining leukofilter-derived CD34+ hematopoietic progenitors and their in vitro differentiation and maturation into proplatelet-bearing megakaryocytes that are able to release platelets in the culture medium. This procedure is useful for in-depth analysis of cellular and molecular mechanisms controlling megakaryopoiesis.

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Biology

Proplatelet Formation Dynamics of Mouse Fresh Bone Marrow Explants
Inès Guinard 1, François Lanza 1, Christian Gachet 1, Catherine Léon 1, Anita Eckly 1
1Université de Strasbourg, INSERM, EFS Grand Est, BPPS UMR-S 1255, FMTS

Here, we detail the bone marrow explant method, from sample preparation to microscopic slide analysis, to evaluate the ability of megakaryocytes which have differentiated in their physiological environment to form proplatelets.

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Biology

Megakaryocyte Culture in 3D Methylcellulose-Based Hydrogel to Improve Cell Maturation and Study the Impact of Stiffness and Confinement
Julie Boscher 1, Christian Gachet 1, François Lanza 1, Catherine Léon 1
1Université de Strasbourg, INSERM, EFS Grand Est, BPPS UMR-S 1255, FMTS

It is now acknowledged that the three-dimensional environment of cells can play an important role in their behavior, maturation and/or differentiation. This protocol describes a three-dimensional cell culture model designed to study the impact of physical containment and mechanical constraints on megakaryocytes.

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Biology

In Vivo Two-photon Imaging of Megakaryocytes and Proplatelets in the Mouse Skull Bone Marrow
Alicia Bornert 1, Fabien Pertuy 1, François Lanza 1, Christian Gachet 1, Catherine Léon 1
1INSERM, EFS Grand Est, Université de Strasbourg

We describe here the method for imaging megakaryocytes and proplatelets in the marrow of the skull bone of living mice using two-photon microscopy.

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