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With this culture device, cells are grown on a continuously expanding culture surface area as the cell population grows.
Whereas the yield obtained with adherent cells is generally limited by the cell culture surface area, suspension-grown cell lines show better and more straightforward scalability.
Six-well culture plates were seeded at 2.1 × 104 undifferentiated MSCs per cm2 of tissue culture surface area in 0.3 ml of media per tissue culture surface.
Infection was carried out under appropriate Biosafety Level 3 conditions using approximately 6 X 10 pfu of rickettsiae/cm of culture surface area.
Briefly, cells were lysed in QIAzol reagent (800 μl/10 cm culture surface area), and the lysate vigorously mixed with 160 μl of chloroform before separating the aqueous phase via centrifugation (12 000 × g, 15 min, 4 °C).
The kidney and spleen pool, cloacal swab, and feather pulp suspensions were centrifuged at 800 x g for 30 min at 4°C, and 1 mL of the supernatant was injected onto an established Vero (ATCC CRL-1587) cell monolayer in 12 cm (culture surface area) bottles.
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Consequently, a robust matrix mineralization was achieved, covering >90% of the culturing surface area.
For TUNEL assay SK-N-SH cells were seeded in BD Falcon™ 8-well culture slides (surface area 0.7 cm2/well) at 104 cells/well.
PAECs were grown in 100 mm culture dishes (surface area 78.5 cm/dish) with an estimated 6 million cells/dish at confluence.
Following fabrication, hydrogels were placed in cell culture medium at surface area:fluid volume ratio of 3 cm/mL and incubated for 24 h at 37 °C.
Control-RBCs maintained at the same density as those in malaria culture also lost some surface area but at a lower rate than URBCs co-cultured with malaria parasites.
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