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Temperature and oxygen concentration profiles are calculated and compared to those predicted by the continuous medium transfer model.
Whereas bystander IMR-90 showed a robust RIBE in medium transfer experiments, hMSC exposed to bystander IMR-90 medium failed to do so.
To mechanistically interrogate RIBE responses and to gain novel insights into RIBE specifically in hSC compartment, both medium transfer and cell co-culture bystander protocols were employed.
With the other types of human cells, we observed the magnitude of RIBE to be no less than 2-fold over sham-exposed cells with bystander medium transfer technique [14].
We and others reported that the RIBE in human adult differentiated somatic cells can be readily observed both with medium transfer and cell co-culture protocols [5], [13], [14].
For example, the magnitude of RIBE for DDR activation in WI38 human fibroblasts was 2.5-fold with medium transfer, about 3-fold with CMRA-staining cell co-culture studies, and 3.7-fold with α-particle IR exposures [5].
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Constructs were cultured in chondrogenic medium for 21 days and thereafter were either maintained in chondrogenic medium, transferred to hypertrophic medium, or implanted subcutaneously into nude mice.
Then, each cell pellet was suspended in fresh Postgate E medium (transferred with a syringe into the Vacutainer tube) and incubated for 7 days at 32°C.
Subsequently, the cells were resuspended in 10-mL cell culture medium, transferred to a 50-mL conical centrifugation tube (Greiner Bio-One) and centrifuged at 200g (1000 rpm) for 5 min.
Cells were washed in DMEM/F12 medium, transferred to RLT lysis buffer (Quiagen) and mercaptoethanol 0.1 µl/ml.
The worms were washed off the plates using screen medium, transferred to two separate beakers, and re-suspended at a density of 1 2 worm/µl in screen medium.
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