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Previous work has produced approaches of varying yield and complexity but does not permit frozen storage of cells for subsequent differentiation in culture.
As compared to static seeding, dynamic cell seeding in spinner flasks resulted in equal cell viability and proliferation, and better cell distribution throughout the scaffold as visualized by histology and confocal microscopy, and was, therefore, selected for subsequent differentiation studies.
Note that this result does not mean that transcriptional activity is unnecessary for subsequent differentiation processes, given that we explored, in this study, a differentiation process that occurs within 24 h of the initial NGF stimulation.
Interestingly, this type of muscle and tendon interaction also operates in Drosophila, where tendon precursor cells are specified in the absence of muscles, but muscle attachment is required for subsequent differentiation of Drosophila tendon cells [74], [75].
Faulty reprogramming at the blastocyst stage would have critical consequences for subsequent differentiation and tissue organization, resulting in large-scale embryo loss at around the implantation stage [1], [19].
The initial differences that would be necessary in order for subsequent differentiation between neighbouring supercolonies to occur are likely to accumulate in allopatry, e.g. if supercolonies stem from different source populations or if supercolonies are divided into two or more fragments through human-mediated jump dispersal.
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For example, the transcription factor Sip1 (Zeb2) is required for the separation of lens progenitors from corneal precursors as well as for the subsequent differentiation of secondary fibers: deletion of Sip1 in the presumptive lens ectoderm led to the persistence of a lens stalk (Yoshimoto et al., 2005).
Further, it has been proposed that down-regulation of Rest in NPCs is required for their subsequent differentiation [8].
Adhesion is an early step in the recruitment of monocytes to inflammatory foci and a prerequisite for their subsequent differentiation to macrophages.
Amplification of a portion of the gene encoding the rickettsial-common 17-kDa antigen allowed for identification and subsequent differentiation by restriction fragment length polymorphism of both Rickettsia typhi and R. felis in C. felis [22].
Thus, our results might help to guide the timely application of these factors for the expansion and subsequent differentiation of osteoblastic cells in culture.
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