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Although 48 hours after cultures became confluent in a 4 cultureture well, significantly more EP bridges (117.3±18.2) formed in EPs grown in BEGM alone than in EPs exposed to FBS in the mixed growth medium (56.0±2.6, Table 1), suggesting FBS does affect EP bridge formation.
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Supernatant containing fibroblasts was centrifuged at 485 g for 8 min; the pellet obtained was suspended in fibroblasts growth medium (Medium 199 and Ham's F12 mixed 1 1 and supplemented with 10% FBS) and cultured at 37°C in 5% CO2.
The sequential substrate uptake observed in mixed substrate growth medium, however, suggest that it is a result of strict CCR.
Thus, the initially simple environment becomes a mixed-substrate growth medium where metabolic interactions can take place.
Half the bacteria were then mixed with more growth medium to keep them alive until they returned to Earth; the other half were mixed with a chemical fixative that stopped their growth and preserved them so that their gene expression could be studied after the shuttle landed.
Mononucleated cells were filtered (40 μm) and plated as a mixed culture in growth medium.
Then the culture medium was replaced with SFM (negative control) or CM mixed with HUVEC growth medium (1 1) with or without Aca1 (10, 25, 50 nM) and/or SU1498 5 μM.
Cells (10) were mixed thoroughly with growth medium (Methocult™ CFU-GM, StemCell Technologies, London, UK), and incubated at 37°C for 7 days in a humidified chamber with 5% CO2.
The precipitates were mixed in (defined keratinocyte growth medium) DKGM to form cell suspension.
About 2.5×10 cells were mixed with 3 ml growth medium containing 0.4% agarose and layered onto 2 ml of 0.75% agarose/medium beds in 6-well plates.
HUVEC were cultured for 24 h on collagen I in presence of CM from LN18 and LN229 cells mixed 1 1 with HUVEC growth medium.
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