2D Co-culture of Leukemic Cells with BMSCs: A Method to Obtain Subpopulations of Leukemic Cells
Published: April 30, 2023
Abstract
Source: Slone, W. L. et al. Modeling Chemotherapy Resistant Leukemia In Vitro. J. Vis. Exp. (2016).
This video describes the technique for long term 2D co-culture of leukemia cells with bone marrow stromal cells to study their interaction with the bone marrow microenvironment. The 2D coculture model utilizes the acute lymphoblastic leukemia cells and the bone marrow stromal or osteoblasts that influence tumor cell phenotype. This study will support investigation of the mechanisms that underlie bone marrow supported survival of leukemic cells during chemotherapy exposure.
Protocol
1. Advanced Preparation Culturing bone marrow stromal cells (BMSC) and osteoblasts (OB). Maintain both BMSC or OB at 37 °C in 6% CO2 and grown on 10 cm tissue culture plates until 90% confluency is reached. Trypsinize BMSC or OB cells and split 1:2 onto new 10 cm plates. The cells are grown to these standards until needed for leukemic co-culturing. 2. Establishing and Maintaining Co-culture Add 5-20 x 10<s…
開示
The authors have nothing to disclose.
Materials
| 100 x 20 mm Cell Culture Dishes | Greiner Bio-One | 664160 | |
| 15 ml conical centrifuge tubes | World Wide Medical Products | 41021037 | Used for cell collection |
| Culture Media | |||
| Osteoblast culture media | PromoCell | C-27001 | For human osteoblast media |
| RPMI 1640 media | Mediatech, Inc. | 15-040 | For tumor media prepation |
| Adherent cells | |||
| Human Osteoblasts | PromoCell | C-12720 | Human osteoblast were cultured according to the supplier’s recommendations. |
| Human Bone Morrow Stromal Cells | WVU Biospecimen Core | De-identified primary human leukemia and bone marrow stromal cells (BMSC) were provided by the Mary Babb Randolph Cancer Center (MBRCC) Biospecimen Processing Core and the West Virginia University Department of Pathology Tissue Bank | |
| Leukemic Cells | |||
| REH | ATCC | ATCC-CRL-8286 | REH cells were cultured according to the supplier’s recommendations and recommended media. |
| SD-1 | DSMZ | ACC 366 | SD-1 were cultured according to the supplier’s recommendations and recommended media. |
