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…

Açıklamalar

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.

Play Video

Bu Makaleden Alıntı Yapın
2D Co-culture of Leukemic Cells with BMSCs: A Method to Obtain Subpopulations of Leukemic Cells. J. Vis. Exp. (Pending Publication), e20260, doi: (2023).

View Video