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An alternative sampling approach targets high-passage events such as diurnal peaks or periods when total daily upstream escapement exceeds 25 000 fish d−1, for increased sampling effort while reducing sampling effort during low-passage periods.
In preliminary experiments, we found that rat AF cells express similar levels of types I and II collagen, sox9, and aggrecan during low-passage culture.
During this study, cell line batches from low passage (< 15) and high passages (> 15) were used.
Contigs or parts thereof consisting only of low passage DNA derived clones were considered to have been lost during cultivation.
Using RNA obtained from low passage (young) and late passage (senescent) fibroblasts, we confirmed that the expression of the mRNA encoding for p16INK4a increased during senescence, as expected.
All experiments were carried out with cultures of low passage (passage number four to eight).
The cells were cultured at low passage (passage number 6 to 8).
All cell lines were maintained in low passage (<15).
At low passage numbers, HPV-immortalized cells are non-tumorigenic.
Immunohistochemical expression analysis of the different key metabolic proteins was also performed using epithelial organotypic rafts cultures obtained from low passage keratinocytes expressing HPV16 E6 and/or E7 expression, as well as from PHK transfected with HPV16 or 18 full-length genomes and grown during different passages before raft culture seeding.
As organotypic cultures obtained from HPV-positive cells at low passage number (FK16A p19 and FK18B p27) exhibit morphological alterations reminiscent of mild/moderate dysplasia while those obtained from the same cells at high passage number (p > 80) exhibit more severe dysplastic features, we can hypothesise that MCT4, CD147, GLUT1 and CAIX are increased during HPV-associated lesion.
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