Dynabeads™ Untouched™ Human B Cells Kit

Protocol tips

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	HCL and HCLv cells for microarray studies were obtained in sodium heparin tubes and purified by Ficoll centrifugation, followed by total B-cell isolation using the Dynabeads™ Untouched™ Human B Cells Kit (ThermoFisher Scientific).

Protocol tips
HCL and HCLv cells for microarray studies were obtained in sodium heparin tubes and purified by Ficoll centrifugation, followed by total B-cell isolation using the Dynabeads™ Untouched™ Human B Cells Kit (ThermoFisher Scientific).

Publication protocol

Samples from patients with HCL, HCLv and other leukemias or controls were obtained via protocol 10-C-0066 approved by the National Cancer Institute Institutional Review Board. Patients gave written informed consent. All patients were adults and therefore did not require consent from parents or guardians. Per protocol, consent is waived in cases where patients died. Patients were consented from March 2010 to August 2019. Patients consented were either inquiring about clinical trials or had questions related to their disease and were invited to give consent to submit research samples. Demographic details of the patients are shown in Table 1. Patients included had a diagnosis of a hematologic malignancy, regardless of the status of their disease. The protocol also allowed recruitment of normal controls. Most of the samples were from patients with HCL and HCLv. The patient population was skewed toward those patients with multiply relapsed disease, although patients with newly diagnosed HCL were also consented. HCL and HCLv cells for microarray studies were obtained in sodium heparin tubes and purified by Ficoll centrifugation, followed by total B-cell isolation using the Dynabeads™ Untouched™ Human B Cells Kit (ThermoFisher Scientific). This procedure removes cells binding to CD2, CD14, CD16a, CD16b, CD36, CD43 and CD235a, including human T cells, monocytes, NK cells, macrophages, granulocytes, plasma cells, platelets, and erythrocytes (per the manual). The HCL and HCLv samples were >80% pure prior to microarray studies.

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