High-throughput enrichment and isolation of megakaryocyte progenitor cells from the mouse bone marrow

Bush, Lucas M.; Healy, Connor P.; Marvin, James; Deans, Tara L.

doi:10.1038/s41598-021-87681-2

Cited by 8 publications

(5 citation statements)

References 41 publications

(26 reference statements)

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“…Bone marrow dissection was performed as previously described [ 20 ]. Briefly, mice were euthanized by CO 2 asphyxiation, followed by cervical dislocation.…”

Section: Methodsmentioning

confidence: 99%

A Comparison of Lymphoid and Myeloid Cells Derived from Human Hematopoietic Stem Cells Xenografted into NOD-Derived Mouse Strains

et al. 2023

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Humanized mice are an invaluable tool for investigating human diseases such as cancer, infectious diseases, and graft-versus-host disease (GvHD). However, it is crucial to understand the strengths and limitations of humanized mice and select the most appropriate model. In this study, we describe the development of the human lymphoid and myeloid lineages using a flow cytometric analysis in four humanized mouse models derived from NOD mice xenotransplanted with CD34+ fetal cord blood from a single donor. Our results showed that all murine strains sustained human immune cells within a proinflammatory environment induced by GvHD. However, the Hu-SGM3 model consistently generated higher numbers of human T cells, monocytes, dendritic cells, mast cells, and megakaryocytes, and a low number of circulating platelets showing an activated profile when compared with the other murine strains. The hu-NOG-EXL model had a similar cell development profile but a higher number of circulating platelets with an inactivated state, and the hu-NSG and hu-NCG developed low frequencies of immune cells compared with the other models. Interestingly, only the hu-SGM3 and hu-EXL models developed mast cells. In conclusion, our findings highlight the importance of selecting the appropriate humanized mouse model for specific research questions, considering the strengths and limitations of each model and the immune cell populations of interest.

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“…Bone marrow dissection was performed as previously described [ 20 ]. Briefly, mice were euthanized by CO 2 asphyxiation, followed by cervical dislocation.…”

Section: Methodsmentioning

confidence: 99%

A Comparison of Lymphoid and Myeloid Cells Derived from Human Hematopoietic Stem Cells Xenografted into NOD-Derived Mouse Strains

et al. 2023

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“…The use of imaging flow cytometry is another way to differentiate MKs from leukocyte–platelet aggregates ( 114 ). It also adds morphological information using the shape, density, texture, and coincidence of fluorescence to discriminate different MK maturation steps ( 115 ). Morphology is an important parameter of MKs that can be obtained by immunohistology or immunofluorescence.…”

Section: Mk Identity and Methodology Advices For Further Studiesmentioning

confidence: 99%

Occurrence and role of lung megakaryocytes in infection and inflammation

2022

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Megakaryocytes (MKs) are large cells giving rise to platelets. It is well established that in adults, MKs develop from hematopoietic stem cells and reside in the bone marrow. MKs are also rare but normal constituents of the venous blood returning to the lungs, and MKs are found in the lung vasculature (MKcirc), suggesting that these cells are migrants from the bone marrow and get trapped in lung capillaries where the final steps of platelet production can occur. An unprecedented increase in the number of lung and circulating MKs was described in coronavirus disease 2019 (COVID-19) patients, suggesting that lung thrombopoiesis may be increased during lung infection and/or thromboinflammation. In addition to the population of platelet-producing intravascular MKs in the lung, a population of lung-resident megakaryocytes (MKL) has been identified and presents a specific immune signature compared to its bone marrow counterparts. Recent single-cell analysis and intravital imaging have helped us gain a better understanding of these populations in mouse and human. This review aims at summarizing the recent data on increased occurrence of lung MKs and discusses their origin, specificities, and potential role in homeostasis and inflammatory and infectious lung diseases. Here, we address remaining questions, controversies, and methodologic challenges for further studies of both MKcirc and MKL.

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“…Immunohistochemistry analysis of major thrombopoiesis organs (bone marrow and spleen) in mice revealed that PALLD deficiency had no effect on the number of megakaryocytes (Figure 3A-B). 28,29 CFU-MK assay indicated that PALLD deficiency did not impact the ability of bone marrow and fetal liver-derived cells to form colonies (Figure 3C). 30 The plasma TPO remained at a similar level in PALLD f/f and PALLD -/mice (Figure 3D).…”

Section: Palld Deficiency Impaired Megakaryocyte Differentiationmentioning

confidence: 99%

The role of PALLD-STAT3 interaction in megakaryocyte differentiation and thrombocytopenia treatment

Li,

Jiang,

Wang

et al. 2024

haematol

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Impaired differentiation of megakaryocytes constitutes the principal etiology of thrombocytopenia. The signal transducer and activator of transcription 3 (STAT3) is a crucial transcription factor in regulating megakaryocyte differentiation, yet the precise mechanism of its activation remains unclear. PALLD, an actin-associated protein, has been increasingly recognized for its essential functions in multiple biological processes. This study revealed that megakaryocyte/plateletspecific knockout of PALLD in mice exhibited thrombocytopenia due to diminished platelet biogenesis. In megakaryocytes, PALLD deficiency led to impaired proplatelet formation and polyploidization, ultimately weakening their differentiation for platelet production. Mechanistic studies demonstrated that PALLD bound to STAT3 and interacted with its DNA-binding domain (DBD) and Src homology 2 (SH2) domain via Immunoglobulin domain 3 (Ig3). Moreover, the absence of PALLD attenuated STAT3 Y705 phosphorylation and impeded STAT3 nuclear translocation. Based on the PALLD-STAT3 binding sequence, we designed a peptide C-P3, which can facilitate megakaryocyte differentiation and accelerate platelet production in vivo. In conclusion, this study highlights the pivotal role of PALLD in megakaryocyte differentiation and proposes a novel approach for treating thrombocytopenia by targeting the PALLD-STAT3 interaction.

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High-throughput enrichment and isolation of megakaryocyte progenitor cells from the mouse bone marrow

Cited by 8 publications

References 41 publications

A Comparison of Lymphoid and Myeloid Cells Derived from Human Hematopoietic Stem Cells Xenografted into NOD-Derived Mouse Strains

A Comparison of Lymphoid and Myeloid Cells Derived from Human Hematopoietic Stem Cells Xenografted into NOD-Derived Mouse Strains

Occurrence and role of lung megakaryocytes in infection and inflammation

The role of PALLD-STAT3 interaction in megakaryocyte differentiation and thrombocytopenia treatment

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