Cryopreservation of cord blood CD34+ cells before or after thrombopoietin expansion differentially affects early platelet recovery in NOD SCID mice

Hensbergen, Yvette van; Garde, Mark van der; Brand, Anneke; Slot, Manon C.; Graaf-Dijkstra, Alice de; Watt, Suzanne M.; Zwaginga, Jaap Jan

doi:10.1111/trf.13045

Cited by 3 publications

(4 citation statements)

References 27 publications

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“…The NOD/SCID mouse model is a very valuable and increasingly used tool for in vivo assessment of human PLT survival. Its application ranges from antibody studies to the in vivo assessment of PLT production and function . Given its potential for transfusion medicine and immunohematology research, it would be highly desirable to make the methods of the model more accessible for the researcher in the field.…”

Section: Discussionmentioning

confidence: 99%

Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations

et al. 2016

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Section: Discussionmentioning

confidence: 99%

Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations

et al. 2016

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“…Cryopreservation has the potential to induce phenotypic changes and can drastically decrease cell viability. Changes induced by cryopreservation have been explored for Bcells [5][6][7][8] , T-cells 9,10 and other hematopoietic cell sub-populations 11,12 . Therefore, evaluating the differences between freshly isolated and frozen cells is necessary to understand the potential effects that cryopreservation may have on downstream functional analyses.…”

Section: Introductionmentioning

confidence: 99%

“…Concordance of functional results between fresh and frozen samples was used as a measure of reliability for each media. As distribution of cell maturation can be altered by cryopreservation 11,12 and culturing in cytokine-enriched media, we assayed for alterations in specific cell surface maturation markers using fluorescence-activated cell sorting (FACS).…”

Section: Introductionmentioning

confidence: 99%

Novel Method Enabling the Use of Cryopreserved Primary Acute Myeloid Leukemia Cells in Functional Drug Screens

Degnin

Agarwal

Tarlock

et al. 2017

Journal of Pediatric Hematology/Oncology

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The ability to assess antileukemic drug activity on primary patient samples is a powerful tool in determining potential drug targets and selection of therapeutic agents with biological and functional rationale. We previously established small molecule inhibitor screens for use on freshly isolated leukemia cells for this purpose. Here we describe a method that produces functional small molecule inhibitor screening results using cryopreserved primary acute myeloid leukemia cells. This method was established to take advantage of biorepositories containing archival material, such as those established by the Children's Oncology Group, and to enable validation of potential pathway dependencies uncovered by genomic analysis. Various conditions used to thaw and culture cryopreserved specimens were assessed for effect on viability, differentiation, and the ability to recapitulate sensitivity results obtained on fresh samples. The most reproducible results were obtained by quick-thawing and culturing samples in cytokine rich media before performing drug screens. Our data suggest that cytokine-enriched media aids in maintaining the viability and numbers required to perform functional analysis on cryopreserved leukemia cells. This method can aid in producing informative data on therapeutic targeting and precision medicine efforts in leukemia by making use of biorepositories and bio banks.

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“…We have taken an alternative approach by producing a cryopreservable cell product consisting of MKs at different stages of maturation that would release PLTs following their transfusion into thrombocytopenic patients. Proof‐of‐principle studies have already demonstrated that human MKs derived ex vivo from either peripheral blood (PB) or umbilical cord blood (UCB) CD34+ cells can generate PLTs after infusion into either mice or non‐human primates . Moreover, the results of a recent Phase I clinical trial demonstrated that infusing UCB‐derived MKs into patients with advanced hematological malignancies was well tolerated .…”

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confidence: 99%

Pre‐clinical development of a cryopreservable megakaryocytic cell product capable of sustained platelet production in mice

et al. 2019

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BACKGROUND Platelet (PLT) transfusions are the most effective treatments for patients with thrombocytopenia. The growing demand for PLT transfusion products is compounded by a limited supply due to dependency on volunteer donors, a short shelf‐life, risk of contaminating pathogens, and alloimmunization. This study provides preclinical evidence that a third‐party, cryopreservable source of PLT‐generating cells has the potential to complement presently available PLT transfusion products. STUDY DESIGN AND METHODS CD34+ hematopoietic stem/progenitor cells derived from umbilical cord blood (UCB) units were used in a simple and efficient culture system to generate a cell product consisting of megakaryocytes (MKs) at different stages of development. The cultures thus generated were evaluated ex vivo and in vivo before and after cryopreservation. RESULTS We generated a megakaryocytic cell product that can be cryopreserved without altering its phenotypical and functional capabilities. The infusion of such a product, either fresh or cryopreserved, into immune‐deficient mice led to production of functional human PLTs which were observed within a week after infusion and persisted for 8 weeks, orders of magnitude longer than that observed after the infusion of traditional PLT transfusion products. The sustained human PLT engraftment was accompanied by a robust presence of human cells in the bone marrow (BM), spleen, and lungs of recipient mice. CONCLUSION This is a proof‐of‐principle study demonstrating the creation of a cryopreservable megakaryocytic cell product which releases functional PLTs in vivo. Clinical development of such a product is currently being pursued for the treatment of thrombocytopenia in patients with hematological malignancies.

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Cryopreservation of cord blood CD34+ cells before or after thrombopoietin expansion differentially affects early platelet recovery in NOD SCID mice

Cited by 3 publications

References 27 publications

Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations

Assessment of human platelet survival in the NOD/SCID mouse model: technical considerations

Novel Method Enabling the Use of Cryopreserved Primary Acute Myeloid Leukemia Cells in Functional Drug Screens

Pre‐clinical development of a cryopreservable megakaryocytic cell product capable of sustained platelet production in mice

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