Road blocks in making platelets for transfusion

Thon, Jonathan N.; Medvetz, DA; Karlsson, SM; Italiano, Jr Je

doi:10.1111/jth.12942

Cited by 36 publications

(35 citation statements)

References 78 publications

(112 reference statements)

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“…Our study shows the efficient generation of HLA-universal HLA-deficient iPSCs. However, the same group has later recognized that β2M -/-cells are susceptible targets for NK cell activity and discuss the co-engineering of iPSCs with the HLA-E molecule, which is known to inhibit NK cell activity (40). Based on our studies, we believe that RNAi might be a superior tool to reduce the immunogenicity of iPSC-derived cell products because silencing HLA to upregulate CD62P and to aggregate.…”

Section: Resultsmentioning

confidence: 98%

Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions

Börger

Eicke

Wolf

et al. 2016

Mol Med

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Platelet (PLT) transfusion is indispensable to maintain homeostasis in thrombocytopenic patients. However, PLT transfusion refractoriness is a common life-threatening condition observed in multitransfused patients. The most frequent immune cause for PLT transfusion refractoriness is the presence of alloantibodies specific for human leukocyte antigen (HLA) class I epitopes. Here, we have silenced the expression of HLA class I to generate a stable HLA-universal induced pluripotent stem cell (iPSC) line that can be used as a renewable cell source for the generation of low immunogenic cell products. The expression of HLA class I was silenced by up to 82% and remained stable during iPSC cultivation. In this study, we have focused on the generation of megakaryocytes (MK) and PLTs from a HLA-universal iPSC source under feeder-and xeno-free conditions. On d 19, differentiation rates of MKs and PLTs with means of 58% and 76% were observed, respectively. HLA-universal iPSC-derived MKs showed polyploidy with DNA contents higher than 4n and formed proPLTs. Importantly, differentiated MKs remained silenced for HLA class I expression. HLA-universal MKs produced functional PLTs. Notably, iPSC-derived HLA-universal MKs were capable to escape antibody-mediated complement-and cellular-dependent cytotoxicity. Furthermore, HLA-universal MKs were able to produce PLTs after in vivo transfusion in a mouse model indicating that they might be used as an alternative to PLT transfusion. Thus, in vitro produced low immunogenic MKs and PLTs may become an alternative to PLT donation in PLT-based therapies and an important component in the management of severe alloimmunized patients.

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Section: Resultsmentioning

confidence: 98%

Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions

Börger

Eicke

Wolf

et al. 2016

Mol Med

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“…According to the GMP for pharmaceutical products by the Food and Drug Administration (FDA), the in vitro manufacture of PLTs requires the consideration of following aspects [42]: (1) Safety of the progenitor cell source. The medical history of the stem cell donor should be known [18] or the cell reprogramming strategy of the source for induced pluripotent stem cells (iPSCs) need to comply with GMP. In addition, reprogramming approaches would preferentially use nonintegrative vectors.…”

Section: Mimicking Megakaryopoiesis and Thrombopoiesis In Vitromentioning

confidence: 99%

“…Essentially, in vitro generated MKs need to satisfy the natural characteristics of MKs derived from bone marrow [51] or cord blood [52]. Thon and colleagues [18] suggested to test in vitro MKs for their morphology, ultrastructure, cytoskeletal organization, granule content, biomarker expression, ploidy, gene expression as well as proPLT formation and PLT release. PLT quality has to be appropriate and correspond to morphological as well as functional characteristics of donor PLTs.…”

Section: Mimicking Megakaryopoiesis and Thrombopoiesis In Vitromentioning

confidence: 99%

Towards the Manufacture of Megakaryocytes and Platelets for Clinical Application

Baigger¹,

Blasczyk²,

Figueiredo³

2017

Transfus Med Hemother

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Platelet transfusions are used in standard clinical practice to prevent hemorrhage in patients suffering from thrombocytopenia or platelet dysfunctions. Recently, a constant rise on the demand of platelets for transfusion has been registered. This may be associated with several factors including demographic changes, population aging as well as incidence and prevalence of hematological diseases. In addition, platelet-regenerative properties have been started to be exploited in different areas such as tissue remodeling and anti-cancer therapies. These new applications are also expected to increase the future demand on platelets. Thus, in vitro generated platelets may constitute a highly desirable alternative to meet the rising demand on platelets. Several factors have been considered in the road trip of producing in vitro megakaryocytes and platelets for clinical application. From selection of the cell source, differentiation protocols and culture conditions to the design of optimal bioreactors, several strategies have been proposed to maximize production yields while preserving functionality. This review summarizes new advances in megakaryocyte and platelet differentiation and their production upscaling.

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“…In vitro MKs or PLTs have to match the phenotype, morphology and ultrastructure of their natural paragons [7]. Importantly, appropriate in vivo models and lately clinical studies need to demonstrate the functional capacity of MKs to produce PLTs in the recipient's circulation.…”

mentioning

confidence: 99%

Large-Scale Cultures and Bioreactors for the Production of Megakaryocytes and Platelets

Baigger¹,

Eicke²,

Blasczyk³

et al. 2017

Adv Tech Biol Med

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A constant rising demand on platelets has been observed which is associated with population ageing and rapid progresses in the development of advanced therapeutic strategies. Therefore, innovative attempts towards the in vitro production of platelets and their precursors are inevitable. Extensive research has been performed on the exploration of reliable cell sources and the establishment of efficient protocols for the differentiation of functional platelets. In particular, the design of bioreactors mimicking the bone marrow microenvironment has gained plenty of attention to achieve clinically relevant platelet yields. In this review, we summarize major advances and perspectives on the manufacture and application of in vitro generated megakaryocytes and platelets.

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Road blocks in making platelets for transfusion

Cited by 36 publications

References 78 publications

Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions

Generation of HLA-Universal iPSC-Derived Megakaryocytes and Platelets for Survival Under Refractoriness Conditions

Towards the Manufacture of Megakaryocytes and Platelets for Clinical Application

Large-Scale Cultures and Bioreactors for the Production of Megakaryocytes and Platelets

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