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MB Sample ID: SA273982
| Local Sample ID: | 52 |
| Subject ID: | SU002832 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Collection:
| Collection ID: | CO002825 |
| Collection Summary: | The spleen organ tissue were harvested, finely minced, and each organ placed individually in 2mL Eppendorf tubes containing 1mL of DMEM with Collagenase D (Sigma Aldrich, product #11088866001). The tissues were incubated at 37C with agitation for 30 minutes. After incubation, 100 uL of 0.1M EDTA was added to the tissue-containing tubes and placed on ice. A single cell suspension was created by addition of Hanks buffered salt solution (HBSS, Corning, product #MT21022CV), passing through a 100 um filter, and collected in a 15 mL conical tube. The cell suspension was spun at 500g for 5 minutes and the supernatant discarded. The remaining tissue/cell solution was resuspended in 5ml of RBC lysis buffer (Invitrogen eBiosciences, product #00-4333-57), incubated at room temperature for 15 minutes, and centrifuged at 500g for 5 minutes, this step was repeated if RBC presence was sustained. Next, the cells were washed with Miltenyi buffer (HBSS, 0.5M EDTA, Fetal bovine serum), resuspended in 180uL Miltenyi buffer, and incubated on ice with CD11b Microbeads (Miltenyi Biotech, product #130-093-636) for 15minutes. Positive cells were isolated using magnetic LS columns (Miltenyi Biotech, product #130-042-401), collected in 2 mL Eppendorf tubes, counted, and then final pellet aliquots were frozen in liquid nitrogen and stored at -80C. continuously using LabScribe2 and analyzed offline. |
| Sample Type: | Macrophages |
