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Briefly, bone marrow was extracted from mouse femurs and tibias.
Mouse femurs and tibias were dissected out and crushed with a pestle.
Bone marrow cells were extracted from mouse femurs and tibiae by flushing.
In brief, bone marrow cells from mouse femurs and tibias were flushed with RPMI 1640 and cultured in RPMI 1640 with 10% FCS, 50 µM of 2-mercaptoethanol and 20 ng/mL of IL3 (PeproTech, Rocky Hill, NJ).
For preparation of primary macrophages, bone marrow from pre-diabetic, 8 12 week old female NOD mice was prepared by flushing mouse femurs and tibias with ice-cold DMEM supplemented with L-glutamine.
Bone marrow derived macrophages were harvested from 6- to 8-week-old mouse femurs and cultured for 6 days in DMEM supplemented with L-glutamine, non-essential amino acids, 10% fetal bovine serum, and 20% L929 conditioned media.
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We also performed biomechanical testing on mouse femur and fifth vertebra.
In mouse femur and rabbit radii fracture models, local application of slow-release VEGF improved callus calcification and volume [ 63].
Briefly, bone marrow cells were collected from the SKG mouse femur, and 1.0 × 106 bone marrow cells were cultured in a 24-well plate with RPMI-1640 supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, 100 μM 2-ME, and 50 ng/ml recombinant murine granulocyte macrophage colony-stimulating factor.
Indeed, the 0.21 fmol/min/mg specific D2 activity determined in mature primary murine osteoblasts in the current studies is in close agreement with the levels of well-characterized specific D2 activity (0.2 0.3 fmol/min/mg) observed previously in mouse femur and bone marrow [30].
MSCs infused into mice femurs and retrieved at different days expressed genes that encoded predicted factors that play a role in cell differentiation and migration.
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