Density Gradient Centrifugation: A Method to Isolate CLL Cells from Peripheral Blood
Published: April 30, 2023
Abstract
Source: Hay, J. et al. Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking. J. Vis. Exp. (2019).
This video describes the technique of isolating chronic lymphocytic leukemia (CLL) cells from peripheral blood. With this protocol, we will isolate CLL cells from human patient blood sample for subsequent efficient generation of nuclear and cytoplasmic fractions.
Protocol
1. Isolation of CLL Cells from Patient Blood Samples Peripheral blood samples from previously consented CLL patients are received from the clinic in EDTA blood collection tubes, accompanied by the white cell count (WCC). Purify the peripheral blood CLL samples according to the WCC. For WCC < 40 x 106 cells/mL, proceed to step 1.1.1; for WCC ≥ 40 x 106 cells/mL, proceed to step 1.1.2. Pour the contents of all the EDTA blood tubes into a 50 mL conical centri…
Açıklamalar
The authors have nothing to disclose.
Materials
| 15 mL Tube | Griener Bio one | 188271 | |
| 1.5 mL microcentrifuge Tubes | Griener Bio one | 616201 | |
| DMSO | Sigma | D2650 | |
| EDTA | Sigma | EDS | |
| Fetal Bovine Serum | Thermofisher | 10500064 | |
| Histopaque1077 density gradient media | Sigma | H8889 | |
| Trypan Blue Solution | Thermofisher | 15250061 | |
| PBS Tablets | Fisher Scientific | BR0014G | |
| RosetteSep Human B Cell Enrichment Cocktail | Stem Cell Technologies | 15064 | |
| Sigma 3-16P | SciQuip | Centrifuge |
Bu Makaleden Alıntı Yapın
Density Gradient Centrifugation: A Method to Isolate CLL Cells from Peripheral Blood. J. Vis. Exp. (Pending Publication), e20265, doi: (2023).