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Biores Open Access. 2014 Jun 1;3(3):110-6. doi: 10.1089/biores.2014.0008.

Sustained Engraftment of Cryopreserved Human Bone Marrow CD34(+) Cells in Young Adult NSG Mice.

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Department of Immunohematology and Blood Transfusion, Leiden University Medical Center , Leiden, The Netherlands .
Central Laboratory Animal Facility, Leiden University Medical Center , Leiden, The Netherlands .
Department of Pediatrics, Leiden University Medical Center , Leiden, The Netherlands . ; Division of Hematology/Oncology, Hospital for Sick Children/University of Toronto , Toronto, Canada .
Department of Pediatrics, Leiden University Medical Center , Leiden, The Netherlands .


Hematopoietic stem cells (HSCs) are defined by their ability to repopulate the bone marrow of myeloablative conditioned and/or (lethally) irradiated recipients. To study the repopulating potential of human HSCs, murine models have been developed that rely on the use of immunodeficient mice that allow engraftment of human cells. The NSG xenograft model has emerged as the current standard for this purpose allowing for engraftment and study of human T cells. Here, we describe adaptations to the original NSG xenograft model that can be readily implemented. These adaptations encompass use of adult mice instead of newborns and a short ex vivo culture. This protocol results in robust and reproducible high levels of lympho-myeloid engraftment. Immunization of recipient mice with relevant antigen resulted in specific antibody formation, showing that both T cells and B cells were functional. In addition, bone marrow cells from primary recipients exhibited repopulating ability following transplantation into secondary recipients. Similar results were obtained with cryopreserved human bone marrow samples, thus circumventing the need for fresh cells and allowing the use of patient derived bio-bank samples. Our findings have implications for use of this model in fundamental stem cell research, immunological studies in vivo and preclinical evaluations for HSC transplantation, expansion, and genetic modification.


human bone marrow; stem cells; thymus; xenograft model

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