scholarly journals Congruence of location-specific transcriptional programs in intestinal organoids during long-term culture

2019 ◽  
Author(s):  
B. van der Hee ◽  
O. Madsen ◽  
H. Smidt ◽  
J.M. Wells
Keyword(s):  
Gene Expression ◽  
Model System ◽  
Transformed Cells ◽  
Stable Model ◽  
Intestinal Development ◽  
Phenotypic Stability ◽  
Intestinal Organoids ◽  
Tissue Of Origin ◽  
Cancer Phenotypes

AbstractThe emergence of intestinal organoids, as a stem cell-based self-renewable model system, has led to many studies on intestinal development and cell-cell signaling. However, potential issues regarding the phenotypic stability and reproducibility of the methodology during culture still needs to be addressed for different organoids. Here we investigated the transcriptomes of intestinal organoids derived from the same pig as well as batch-to-batch variation of organoids derived from different pigs over long-term passage. The set of genes expressed in organoids closely resembled that of the tissue of origin, including location specific functions, for at least 17 passages. Minor differences in gene expression were observed between individual organoid cultures. In contrast, most tissue-specific genes were not expressed in the transformed jejunum cell line IPECJ2, which also showed gene expression consistent with cancer phenotypes. We conclude that intestinal organoids provide a robust and stable model for translational research with clear advantages over transformed cells.

scholarly journals Phenotypic Stability of Transgenic Citrus

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 903G-904 ◽  
Author(s):  
Gloria A. Moore
Keyword(s):  
Gene Expression ◽  
Transgenic Plants ◽  
Transformed Cells ◽  
Gus Gene ◽  
Phenotypic Stability ◽  
Vector Plasmid ◽  
Primary Transformants ◽  
Transgenic Citrus ◽  
Stem Segments

We have produced a number of transgenic citrus plants via Agrobacterium-mediated transformation of seedling stem segments with a vector plasmid containing a β-glucuronidase (GUS) gene. All regenerated green shoots produced in our experiments are assayed histochemically for expression of GUS by cutting a section from the base of the shoot. Many of the shoots express GUS only in sectors, which vary in size from shoot to shoot. Analyses suggest that sectored regenerated shoots are chimeric, consisting of nontransformed cells as well as transformed cells. However, plants derived from shoots with large GUS+ sectors in the original assays do not necessarily contain the GUS gene; conversely, some plants derived from shoots with small sectors appear solidly transformed. Plants that appear solidly transformed have maintained gene expression for up to 5 years. None of the transgenic plants have obviously altered morphologies. It has not been possible to analyze progeny plants because of the long juvenile periods and polyembryony of the primary transformants. However, because citrus is clonally propagated, long-term phenotypic stability of primary transformants is the most important factor in producing useful transgenic plants.

scholarly journals Development and Validation of a Long-Term 3D Glioblastoma Cell Culture in Alginate Microfibers as a Novel Bio-Mimicking Model System for Preclinical Drug Testing

2021 ◽  
Vol 11 (8) ◽  
pp. 1025
Author(s):  
Miodrag Dragoj ◽  
Jasmina Stojkovska ◽  
Tijana Stanković ◽  
Jelena Dinić ◽  
Ana Podolski-Renić ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Culture ◽  
Cell Growth ◽  
Drug Testing ◽  
3D Model ◽  
Model System ◽  
Glioblastoma Cell ◽  
Related Gene ◽  
Alginate Microfibers

Background: Various three-dimensional (3D) glioblastoma cell culture models have a limited duration of viability. Our aim was to develop a long-term 3D glioblastoma model, which is necessary for reliable drug response studies. Methods: Human U87 glioblastoma cells were cultured in alginate microfibers for 28 days. Cell growth, viability, morphology, and aggregation in 3D culture were monitored by fluorescent and confocal microscopy upon calcein-AM/propidium iodide (CAM/PI) staining every seven days. The glioblastoma 3D model was validated using temozolomide (TMZ) treatments 3 days in a row with a recovery period. Cell viability by MTT and resistance-related gene expression (MGMT and ABCB1) by qPCR were assessed after 28 days. The same TMZ treatment schedule was applied in 2D U87 cell culture for comparison purposes. Results: Within a long-term 3D model system in alginate fibers, U87 cells remained viable for up to 28 days. On day 7, cells formed visible aggregates oriented to the microfiber periphery. TMZ treatment reduced cell growth but increased drug resistance-related gene expression. The latter effect was more pronounced in 3D compared to 2D cell culture. Conclusion: Herein, we described a long-term glioblastoma 3D model system that could be particularly helpful for drug testing and treatment optimization.

scholarly journals Agroinfiltration of grapevine leaves for fast transient assays of gene expression and for long-term production of stable transformed cells

2008 ◽  
Vol 27 (5) ◽  
pp. 845-853 ◽  
Author(s):  
Michela Zottini ◽  
Elisabetta Barizza ◽  
Alex Costa ◽  
Elide Formentin ◽  
Cristina Ruberti ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transformed Cells ◽  
Transient Assays ◽  
Grapevine Leaves ◽  
Fast Transient

Long term dietary intake of aromatic amino acids are associated with leptin gene expression in adipose tissues of non-diabetic adults

2018 ◽  
Author(s):  
Golaleh Asghari ◽  
Emad Yuzbashian ◽  
Maryam Zarkesh ◽  
Parvin Mirmiran ◽  
Mehdi Hedayati ◽  
...  
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Dietary Intake ◽  
Aromatic Amino Acids ◽  
Adipose Tissues

Stage 1 Registered Report Manuscript - Transcriptional Correlates of Savings Memory: Is Forgetting Due to Decay or Retrieval Failure?

2020 ◽  
Author(s):  
Robert Calin-Jageman ◽  
Irina Calin-Jageman ◽  
Tania Rosiles ◽  
Melissa Nguyen ◽  
Annette Garcia ◽  
...  
Keyword(s):  
Gene Expression ◽  
Memory Trace ◽  
Aplysia Californica ◽  
Retrieval Failure ◽  
Initial Learning ◽  
Rigorous Test ◽  
Regulated Gene Expression ◽  
Stage 1 ◽  
Weak Similarity

[[This is a Stage 1 Registered Report manuscript. The project was submitted for review to eNeuro. Upon revision and acceptance, this version of the manuscript was pre-registered on the OSF (9/11/2019, https://osf.io/fqh8j) (but due to an oversight not posted as a preprint until July 2020). A Stage 2 manuscript is now posted as a pre-print (https://psyarxiv.com/h59jv) and is under review at eNeuro. A link to the final Stage 2 manuscript will be added when available.]]There is fundamental debate about the nature of forgetting: some have argued that it represents the decay of the memory trace, others that the memory trace persists but becomes inaccessible due to retrieval failure. These different accounts of forgetting make different predictions about savings memory, the rapid re-learning of seemingly forgotten information. If forgetting is due to decay then savings requires re-encoding and should thus involve the same mechanisms as initial learning. If forgetting is due to retrieval-failure then savings should be mechanistically distinct from encoding. In this registered report we conducted a pre-registered and rigorous test between these accounts of forgetting. Specifically, we used microarray to characterize the transcriptional correlates of a new memory (1 day from training), a forgotten memory (8 days from training), and a savings memory (8 days from training but with a reminder on day 7 to evoke a long-term savings memory) for sensitization in Aplysia californica (n = 8 samples/group). We find that the transcriptional correlates of savings are [highly similar / somewhat similar / unique] relative to new (1-day-old) memories. Specifically, savings memory and a new memory share [X] of [Y] regulated transcripts, show [strong / moderate / weak] similarity in sets of regulated transcripts, and show [r] correlation in regulated gene expression, which is [substantially / somewhat / not at all] stronger than at forgetting. Overall, our results suggest that forgetting represents [decay / retrieval-failure / mixed mechanisms].

scholarly journals Malpigmentation of Common Sole (Solea solea) during Metamorphosis Is Associated with Differential Synaptic-Related Gene Expression

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2273
Author(s):  
Menelaos Kavouras ◽  
Emmanouil E. Malandrakis ◽  
Ewout Blom ◽  
Kyriaki Tsilika ◽  
Theodoros Danis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nervous System ◽  
De Novo ◽  
Close Association ◽  
Ionic Channels ◽  
Long Term Potentiation ◽  
General Term ◽  
The Difference ◽  
Common Sole

In farmed flatfish, such as common sole, color disturbances are common. Dyschromia is a general term that includes the color defects on the blind and ocular sides of the fish. The purpose was to examine the difference in gene expression between normal pigmented and juveniles who present ambicoloration. The analysis was carried out with next-generation sequencing techniques and de novo assembly of the transcriptome. Transcripts that showed significant differences (FDR < 0.05) in the expression between the two groups, were related to those of zebrafish (Danio rerio), functionally identified, and classified into categories of the gene ontology. The results revealed that ambicolorated juveniles exhibit a divergent function, mainly of the central nervous system at the synaptic level, as well as the ionic channels. The close association of chromophore cells with the growth of nerve cells and the nervous system was recorded. The pathway, glutamate binding–activation of AMPA and NMDA receptors–long-term stimulation of postsynaptic potential–LTP (long term potentiation)–plasticity of synapses, appears to be affected. In addition, the development of synapses also seems to be affected by the interaction of the LGI (leucine-rich glioma inactivated) protein family with the ADAM (a disintegrin and metalloprotease) ones.

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