scholarly journals Isolation and long-term expansion of murine epidermal stem-like cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0254731
Author(s):  
Jingjing Wang ◽  
Maureen Mongan ◽  
Xiang Zhang ◽  
Ying Xia
Keyword(s):  
Epithelial Cell ◽  
Intercellular Junctions ◽  
Global Gene Expression ◽  
Cell Types ◽  
Proliferation Markers ◽  
Expression Studies ◽  
Sustainable Culture ◽  
Proliferation And Differentiation ◽  
Epidermal Homeostasis ◽  
Gene Expression Studies

Epidermis is the most outer layer of the skin and a physical barrier protecting the internal tissues from mechanical and environmental insults. The basal keratinocytes, which, through proliferation and differentiation, supply diverse cell types for epidermal homeostasis and injury repair. Sustainable culture of murine keratinocyte, however, is a major obstacle. Here we developed murine keratinocyte lines using low-Ca2+ (0.06 mM) keratinocyte serum-free medium (KSFM-Ca2+) without feeder cells. Cells derived in this condition could be subcultured for >70 passages. They displayed basal epithelial cell morphology and expressed keratin (Krt) 14, but lacked the epithelial-characteristic intercellular junctions. Moreover, these cells could be adapted to grow in the Defined-KSFM (DKSFM) media containing 0.15 mM Ca2+, and the adapted cells established tight- and adherens-junctions and exhibited increased Krt1/10 expression while retained subculture capacity. Global gene expression studies showed cells derived in KSFM-Ca2+ media had enriched stem/proliferation markers and cells adapted in DKSFM media had epithelial progenitor signatures. Correspondingly, KSFM-Ca2+-derived cells exhibited a remarkable capacity of clonal expansion, whereas DKSFM-adapted cells could differentiate to suprabasal epithelial cell types in 3-dimentional (3D) organoids. The generation of stem-like murine keratinocyte lines and the conversion of these cells to epithelial progenitors capable of terminal differentiation provide the critically needed resources for skin research.

Deficiency in intercellular communication in two established renal epithelial cell lines (LLC-PK1 and MDCK)

1984 ◽  
Vol 66 (1) ◽  
pp. 81-93
Author(s):  
F.V. Sepulveda ◽  
J.D. Pearson
Keyword(s):  
Epithelial Cell ◽  
Cell Lines ◽  
Mdck Cells ◽  
Cell Communication ◽  
Intercellular Junctions ◽  
Cell Types ◽  
Renal Epithelial Cell ◽  
Aortic Endothelial Cells ◽  
Epithelial Cell Lines ◽  
Autoradiographic Analysis

We have studied the cell-to-cell passage of uridine nucleotides in two renal epithelial cell lines (LLC-PK1 and MDCK) and in porcine aortic endothelial cells (PAE). All three cell types incorporated tritiated uridine. After a 3 h incubation the radioactivity was predominantly in the form of acid-soluble compounds, mainly UTP. Prelabelled LLC-PK1 or MDCK cells were unable to transfer radioactivity to added adjacent, non-labelled cells, whereas PAE cells readily formed communicating intercellular junctions, as judged by autoradiographic analysis after a 3 h co-culture period. Cell-to-cell communication in either of the renal cell lines was not promoted by treatment with dibutyryl cyclic AMP and methylisobutylxanthine. Radioactivity incorporated into the acid-insoluble pool was not available for intercellular transfer, as assessed in experiments in which cells were prelabelled 24 h before co-culture.

scholarly journals Trisomy 21 and Down syndrome: a short review

2008 ◽  
Vol 68 (2) ◽  
pp. 447-452 ◽  
Author(s):  
CA. Sommer ◽  
F. Henrique-Silva
Keyword(s):  
Gene Expression ◽  
Down Syndrome ◽  
Large Scale ◽  
Molecular Mechanisms ◽  
Global Gene Expression ◽  
Short Review ◽  
Complex Disorder ◽  
Expression Studies ◽  
Gene Expression Studies ◽  
Chromosome Segments

Even though the molecular mechanisms underlying the Down syndrome (DS) phenotypes remain obscure, the characterization of the genes and conserved non-genic sequences of HSA21 together with large-scale gene expression studies in DS tissues are enhancing our understanding of this complex disorder. Also, mouse models of DS provide invaluable tools to correlate genes or chromosome segments to specific phenotypes. Here we discuss the possible contribution of HSA21 genes to DS and data from global gene expression studies of trisomic samples.

scholarly journals Mayaro Virus Infects Human Chondrocytes and Induces the Expression of Arthritis-Related Genes Associated with Joint Degradation

Viruses ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 797 ◽  
Author(s):  
Michèle Bengue ◽  
Pauline Ferraris ◽  
Cécile Baronti ◽  
Cheikh Tidiane Diagne ◽  
Loïc Talignani ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Types ◽  
Human Chondrocytes ◽  
Cellular Mechanisms ◽  
Matrix Gene ◽  
Knowing That ◽  
Expression Studies ◽  
Infected Cells ◽  
Gene Expression Studies ◽  
Mayaro Virus

Mayaro virus (MAYV) is an emerging arthritogenic alphavirus belonging to the Togaviridae family. Infection leads to a dengue-like illness accompanied by severe polyarthralgia. However, the molecular and cellular mechanisms of arthritis as a result of MAYV infection remain poorly understood. In the present study, we assess the susceptibility of human chondrocytes (HC), fibroblast-like synoviocytes and osteoblasts that are the major cell types involved in osteoarthritis, to infection with MAYV. We show that these cells are highly permissive to MAYV infection and that viral RNA copy number and viral titers increase over time in infected cells. Knowing that HC are the primary cells in articular cartilage and are essential for maintaining the cartilaginous matrix, gene expression studies were conducted in MAYV-infected primary HC using polymerase chain reaction (PCR) arrays. The infection of the latter cells resulted in an induction in the expression of several matrix metalloproteinases (MMP) including MMP1, MMP7, MMP8, MMP10, MMP13, MMP14 and MMP15 which could be involved in the destruction of articular cartilage. Infected HC were also found to express significantly increased levels of various IFN-stimulated genes and arthritogenic mediators such as TNF-α and IL-6. In conclusion, MAYV-infected primary HC overexpress arthritis-related genes, which may contribute to joint degradation and pathogenesis.

scholarly journals Pathway-Level Information ExtractoR (PLIER) for gene expression data

2017 ◽  
Author(s):  
Weiguang Mao ◽  
Elena Zaslavsky ◽  
Boris M. Hartmann ◽  
Stuart C. Sealfon ◽  
Maria Chikina
Keyword(s):  
Gene Expression ◽  
Global Gene Expression ◽  
Expression Data ◽  
Regulate Gene Expression ◽  
Expression Studies ◽  
Inference Algorithms ◽  
Biological Insight ◽  
Level Information ◽  
Gene Expression Studies ◽  
Cell Proportion

AbstractA major challenge in gene expression analysis is to accurately infer relevant biological insight, such as regulation of cell type proportion or pathways, from global gene expression studies. We present a general solution for this problem that outperforms available cell proportion inference algorithms, and is more widely useful to automatically identify specific pathways that regulate gene expression. Our method improves replicability and biological insight when applied to trans-eQTL identification.

scholarly journals PPIA and YWHAZ Constitute a Stable Pair of Reference Genes during Electrical Stimulation in Mesenchymal Stem Cells

2021 ◽  
Vol 12 (1) ◽  
pp. 153
Author(s):  
Lynsey Steel ◽  
David M. Ansell ◽  
Enrique Amaya ◽  
Sarah H. Cartmell
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Electrical Stimulation ◽  
Reference Gene ◽  
Reference Genes ◽  
Adult Stem Cells ◽  
Expression Studies ◽  
Proliferation And Differentiation ◽  
Gene Expression Studies

Mesenchymal stem cells (MSCs) are multipotent adult stem cells with great potential in regenerative medicine. One method for stimulating proliferation and differentiation of MSCs is via electrical stimulation (ES). A valuable approach for evaluating the response of MSCs to ES is to assess changes in gene expression, relative to one or more reference genes. In a survey of 25 publications that used ES on cells, 70% selected GAPDH as the reference gene. We conducted a study to assess the suitability of six potential reference genes on an immortalized human MSC line following direct current ES at seeding densities of 5000 and 10,000 cells/cm2. We employed three methods to validate the most stable reference genes from qRT-PCR data. Our findings show that GAPDH and ACTB exhibit reduced stability when seeded at 5000 cell/cm2. In contrast, we found that the most stable genes across both plating densities and stimulation regimes were PPIA and YWHAZ. Thus, in ES gene expression studies in MSCs, we support the use of PPIA and YWHAZ as an optimal reference gene pair, and discourage the use of ACTB and GAPDH at lower seeding densities. However, it is strongly recommended that similar verification studies are carried out based on cell type and different ES conditions.

scholarly journals Large-scale gene expression studies of the endometrium: what have we learnt?

Reproduction ◽  
2006 ◽  
Vol 132 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Rob Sherwin ◽  
Rob Catalano ◽  
Andrew Sharkey
Keyword(s):  
Large Scale ◽  
Dna Microarrays ◽  
Disease States ◽  
Complex Interactions ◽  
Expression Studies ◽  
Proliferation And Differentiation ◽  
Potential Applications ◽  
Underlying Causes ◽  
Gene Expression Studies ◽  
The Impact

The endometrium is a dynamic tissue that undergoes coordinated changes under the influence of steroid hormones. This results in proliferation and differentiation culminating in a receptive state, followed by menstruation and endometrial repair. These functions involve complex interactions between the epithelium, stroma and leucocytes in the endometrium. Understanding the underlying causes of endometrial disorders, such as infertility, endometriosis and heavy menstrual bleeding, therefore represents a considerable challenge. Recently developed techniques, such as differential display and DNA microarrays permit the abundance of thousands of mRNA transcripts within cells or tissues to be measured simultaneously. This provides a new approach to understanding the complex interactions that underlie both healthy and disease states. Responses of the endometrium to hormones or drugs can be studied and the response of the system as an integrated whole can be assessed. Comparisons of endometrium from healthy women and those with endometrial dysfunction have advanced our understanding of key areas of endometrial physiology, including infertility, receptivity, endometriosis and cancer. Using this approach, novel genes controlling specific endometrial functions like receptivity have been identified for functional testing. This paper will review the impact of these techniques for transcript profiling on our understanding of selected areas of endometrial biology and discuss the potential applications in future.

Metals and methylotrophy: Via global gene expression studies

2021 ◽  
Author(s):  
Zachary D. Johnson ◽  
Dennis D. Krutkin ◽  
Pavlo Bohutskyi ◽  
Marina G. Kalyuzhnaya
Keyword(s):  
Gene Expression ◽  
Global Gene Expression ◽  
Expression Studies ◽  
Gene Expression Studies
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