Xenotransplantation of human hematopoietic stem cells
Primary hHSPCs (CD34+) cells were purified from the BM or peripheral blood of patients using Ficoll density gradient centrifugation and further magnetically isolated using the MACS CD34 MicroBead Kit (Miltenyi Biotec). CD34+ cells were cryopreserved and slowly thawed in IMDM with 50% FCS at 37 °C before xenotransplantation and resuspended in 25 µl PBS for injection. MISTRG mice aged 8–12 weeks old59 (link) were irradiated sub-lethally (181 cGy using an X-ray RS-2000 irradiator, Rad Source) and transplanted with a 22-gauge needle (Hamilton Company) intrafemorally with 1.5–3 × 105 CD34+ hHSPCs.
Grockowiak E., Korn C., Rak J., Lysenko V., Hallou A., Panvini F.M., Williams M., Fielding C., Fang Z., Khatib-Massalha E., García-García A., Li J., Khorshed R.A., González-Antón S., Baxter E.J., Kusumbe A., Wilkins B.S., Green A., Simons B.D., Harrison C.N., Green A.R., Lo Celso C., Theocharides A.P, & Méndez-Ferrer S. (2023). Different niches for stem cells carrying the same oncogenic driver affect pathogenesis and therapy response in myeloproliferative neoplasms. Nature Cancer, 4(8), 1193-1209.
BloodCells Density gradient centrifugationFicoll Mice Microbead NeedlePatients X ray Xenotransplantation
Corresponding Organization :
Other organizations :
University of Cambridge, Wellcome/MRC Cambridge Stem Cell Institute, Medical Research Council, National Health Service, University Hospital of Zurich, University of Zurich, Wellcome/Cancer Research UK Gurdon Institute, Wellcome Trust, Imperial College London, The Francis Crick Institute, University of Oxford, Guy's and St Thomas' NHS Foundation Trust
Purification of primary hHSPCs (CD34+ cells) from the BM or peripheral blood of patients using Ficoll density gradient centrifugation and further magnetic isolation using the MACS CD34 MicroBead Kit
Cryopreservation and slow thawing of CD34+ cells in IMDM with 50% FCS at 37 °C before xenotransplantation
Irradiation of MISTRG mice aged 8–12 weeks sub-lethally (181 cGy) using an X-ray RS-2000 irradiator
Intrafemorally transplanting 1.5–3 × 10^5 CD34+ hHSPCs into MISTRG mice using a 22-gauge needle
dependent variables
Not explicitly mentioned
control variables
Age of MISTRG mice (8–12 weeks)
positive controls
Not specified
negative controls
Not specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
