In vitro generation of platelets through direct conversion: first report in My Knowledge (iMK)

Masuda, Shigeo; Li, Mo; Belmonte, Juan Carlos Izpisúa

doi:10.1038/cr.2012.142

Cited by 9 publications

(7 citation statements)

References 15 publications

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“…Mounting evidence that cell-cell contacts, 22,23 ECM composition 19,20 and stiffness, 21 vascular shear stress, 24,25 pO2/pH, 51,52 soluble factor interactions, 22,53 and temperature 54 contribute to proPLT formation and PLT release have suggested that recapitulating key components of BM and blood vessel microenvironments within a three-dimensional microfluidic culture system is necessary to achieve clinically significant numbers of functional human PLTs. Our microfluidic bioreactor design expands control and resolution of the microenvironment, and has allowed us to drastically improve the time to PLT release (from 18 hours down to 2 hours) and more For personal use only.…”

Section: Discussionmentioning

confidence: 99%

Platelet bioreactor-on-a-chip

Thon

Mažutis

et al. 2014

Blood

186

210

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• We have developed a biomimetic microfluidic platelet bioreactor that recapitulates bone marrow and blood vessel microenvironments.• Application of shear stress in this bioreactor triggers physiological proplatelet production, and platelet release.Platelet transfusions total >2.17 million apheresis-equivalent units per year in the United States and are derived entirely from human donors, despite clinically significant immunogenicity, associated risk of sepsis, and inventory shortages due to high demand and 5-day shelf life. To take advantage of known physiological drivers of thrombopoiesis, we have developed a microfluidic human platelet bioreactor that recapitulates bone marrow stiffness, extracellular matrix composition, micro-channel size, hemodynamic vascular shear stress, and endothelial cell contacts, and it supports high-resolution live-cell microscopy and quantification of platelet production. Physiological shear stresses triggered proplatelet initiation, reproduced ex vivo bone marrow proplatelet production, and generated functional platelets. Modeling human bone marrow composition and hemodynamics in vitro obviates risks associated with platelet procurement and storage to help meet growing transfusion needs. (Blood. 2014;124(12):1857-1867) IntroductionAlthough platelets (PLTs) play critical roles in hemostasis, 1 angiogenesis, 2 and innate immunity, 3 PLT production remains poorly understood. Consequently, PLT units are derived entirely from human donors, despite serious clinical concerns owing to their immunogenicity and associated risk of sepsis. 4 More than 2.17 million apheresisequivalent PLT units are transfused yearly in the United States 5,6 at a cost of .$1 billion per year. Although demand for PLT transfusions has increased markedly in the past decade, a near-static pool of donors and a 5-day PLT unit shelf life resulting from bacterial contamination 7 and storage-related PLT deterioration, 8 have resulted in significant PLT shortages. 9 Furthermore, artificial platelet substitutes have failed to replace physiological platelet products. 10 An efficient, donorindependent PLT bioreactor capable of generating clinically significant numbers of functional human PLTs is necessary to obviate risks associated with PLT procurement and storage, and help meet growing transfusion needs. In vivo, megakaryocytes (MKs) PLT progenitors sit outside blood vessels in the bone marrow (BM) and extend long, branching cellular structures designated proPLTs into the circulation from which PLTs are released. 11-15 Nearly 100% of human adult MKs must produce ;10 3 PLTs each to account for circulating PLT counts. 16 Although functional human PLTs were first grown in vitro in 1995, 17 to date only ;10% of human MKs initiate proPLT production in culture. This results in yields of 10 122 PLTs per CD34 1 cord blood-derived or embryonic stem cell-derived MK, 18 which are themselves of limited availability, constituting a significant bottleneck in the ex vivo production of a PLT transfusion unit. Although second-generation c...

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Section: Discussionmentioning

confidence: 99%

Platelet bioreactor-on-a-chip

Thon

Mažutis

et al. 2014

Blood

186

210

View full text Add to dashboard Cite

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“…Of course, many other different kinds of cell sources have been found and cultured to generate MKs/PLTs in vitro, such as human embryonic stem cells (hESCs) (Olsen et al 2006 ; Pick et al 2013 ), hematopoietic stem cells (HSCs) (Debili et al 1995 ; Mercher et al 2008 ), induced megakaryocytes (iMKs) (Masuda et al 2012 ), human CD34 + cells from bone marrow (BM), umbilical cord blood (CB) (Choi et al 1995 ; Matsunaga et al 2006 ), and even human endometrial stromal stem cells (hESSCs) (Wang et al 2012 ). However, these cells cannot be utilized satisfactorily due to lack of resources and other reasons; therefore, it is not necessary to describe them in detail in the present article (Table 1 ).…”

Section: Platelet Generation In Vitromentioning

confidence: 99%

Platelet generation in vivo and in vitro

Wang

Zheng

2016

SpringerPlus

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Platelet (PLT) transfusion, which is the primary cell therapy for thrombocytopenia, has been a source of concern in recent years due to its limitations of donor-dependent supply and soaring costs. In vitro platelet generation on an industrial scale is a possible solution requiring exploration. The technology of platelet generation ex vivo has been widely studied across the world, though the mechanisms of physiological thrombopoiesis and platelet biology function in vivo still remain elusive today. Various culture systems have been studied, most of which proved quite inefficient in generating functional platelets ex vivo, so there is still a long way to reach our ultimate goal of generating a fully functional platelet in vitro on an industrial scale. This review integrates the latest research into physiological platelet biogenesis and ex vivo-platelet/megakaryocyte (MK) generation protocols with a focus on the ability to generate PLT/MK in large quantities, summarizes current culture systems based on induced human pluripotent stem cells and adipose-derived stem cells, and discusses significant challenges that must be overcome for these approaches to be perfected.

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“…The alternative approach to eliminate donor dependency and alloimmunization risk is the infusion of ex vivo differentiated MKPs or platelets generated from the patient’s own cells 86–88 . Current studies are investigating the best source of stem cells, timeline and order of cytokine addition to the culture in order to govern stem cell differentiation to MKs 88,89 . NGF/TrkA signaling can support these culture systems at an early stage where MKP enrichment is desired.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Tropomyosin Receptor Kinase A Signaling Negatively Regulates Megakaryopoiesis and induces Thrombopoiesis

Kizilyer¹,

Singh²,

Singh³

et al. 2019

Sci Rep

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Neurotrophin signaling modulates the differentiation and function of mature blood cells. The expression of neurotrophin receptors and ligands by hematopoietic and stromal cells of the bone marrow indicates that neurotrophins have the potential to regulate hematopoietic cell fate decisions. This study investigates the role of neurotrophins and Tropomyosin receptor kinases (Trk) in the development of megakaryocytes (MKs) and their progeny cells, platelets. Results indicate that primary human MKs and MK cells lines, DAMI, Meg-01 and MO7e express TrkA, the primary receptor for Nerve Growth Factor (NGF) signaling. Activation of TrkA by NGF enhances the expansion of human MK progenitors (MKPs) and, to some extent, MKs. Whereas, inhibition of TrkA receptor by K252a leads to a 50% reduction in the number of both MKPs and MKs and is associated with a 3-fold increase in the production of platelets. In order to further confirm the role of TrkA signaling in platelet production, TrkA deficient DAMI cells were generated using CRISPR-Cas9 technology. Comparative analysis of wild-type and TrkA-deficient Dami cells revealed that loss of TrkA signaling induced apoptosis of MKs and increased platelet production. Overall, these findings support a novel role for TrkA signaling in platelet production and highlight its potential as therapeutic target for Thrombocytopenia.

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In vitro generation of platelets through direct conversion: first report in My Knowledge (iMK)

Cited by 9 publications

References 15 publications

Platelet bioreactor-on-a-chip

Platelet bioreactor-on-a-chip

Platelet generation in vivo and in vitro

Inhibition of Tropomyosin Receptor Kinase A Signaling Negatively Regulates Megakaryopoiesis and induces Thrombopoiesis

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