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MB Sample ID: SA237893
| Local Sample ID: | Blank-3 |
| Subject ID: | SU002472 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Subject Comments: | Human B-ALL cell line (REH and RCH-AcV) were a generous gift from Dr. Christopher Porter at Emory University |
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Treatment:
| Treatment ID: | TR002484 |
| Treatment Summary: | REH and RCH-AcV were maintained in RPMI1640 (cat#10-041-CV, Corning, Glendale, AZ), supplemented with 20% FBS (cat#S11550, BioTechne, Minneapolis, MN). OP-9 bone marrow stromal cells were maintained in α-MEM (cat#15-012-CV, Corning, Glendale, AZ) supplemented with 20% FBS. For adipocyte differentiation, 105 OP-9 cells were plated in 6-well plates in DMEM (cat#10-017-CV, Corning, Glendale, AZ) supplemented with 10% FBS as previously described [18, 26]. After 24 hours of culture, the media was removed and switched to differentiation media which is composed of α-MEM supplemented with 1.8mM oleate (cat#O7501, Sigma, St. Louis, MO) bound to BSA (cat#A6003, Sigma, St. Louis, MO) with molar ratio 5.5:1 along with 175nM insulin (cat#I6634, Sigma, St. Louis, MO) and 0.2% FBS. ACM was collected after 3 days of differentiation and used in the experiments describe in this study. For SCM, OP-9 cells were plated in DMEM supplemented with 10% FBS and conditioned media were collected on day 3 of culture. Human B-ALL cell lines (REH and RCH-AcV) were cultured for 24 hours in UCM, SCM, or ACM. Additionally, methotrexate (MTX)-treated human B-ALL cells were cultured for an additional 12 hours of treatment with MTX (50nM for REH and 70 nM for RCH-AcV). This timepoint was chosen due to our prior work in this system to avoid submitting mostly apoptotic cells for analysis. |
| Treatment Compound: | adipocyte- , stromal cell-, and un- conditioned media; methotrexate or not |
| Treatment Dosevolume: | methotrexate: 50-70 nM |
| Treatment Doseduration: | conditioned medias: 24 hr; methotrexate: additional 12 hr |
| Treatment Vehicle: | methotrexate: DMSO |
