scholarly journals A Novel In Vitro Culture Model System to Study Merkel Cell Polyomavirus–Associated MCC Using Three-Dimensional Organotypic Raft Equivalents of Human Skin

Viruses ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 138
Author(s):  
Amanda S. W. Loke ◽  
B. Jack Longley ◽  
Paul F. Lambert ◽  
Megan E. Spurgeon
Keyword(s):  
Culture System ◽  
Three Dimensional ◽  
Human Keratinocytes ◽  
Merkel Cell Polyomavirus ◽  
Three Dimensions ◽  
Human Polyomavirus ◽  
Merkel Cell ◽  
Raft Culture ◽  
Dermal Equivalent

Merkel cell polyomavirus (MCPyV) is a human polyomavirus causally linked to the development of Merkel cell carcinoma (MCC), an aggressive malignancy that largely arises within the dermis of the skin. In this study, we recapitulate the histopathology of human MCC tumors in vitro using an organotypic (raft) culture system that is traditionally used to recapitulate the dermal and epidermal equivalents of skin in three dimensions (3D). In the optimal culture condition, MCPyV+ MCC cells were embedded in collagen between the epidermal equivalent comprising human keratinocytes and a dermal equivalent containing fibroblasts, resulting in MCC-like lesions arising within the dermal equivalent. The presence and organization of MCC cells within these dermal lesions were characterized through biomarker analyses. Interestingly, co-culture of MCPyV+ MCC together with keratinocytes specifically within the epidermal equivalent of the raft did not reproduce human MCC morphology, nor were any keratinocytes necessary for MCC-like lesions to develop in the dermal equivalent. This 3D tissue culture system provides a novel in vitro platform for studying the role of MCPyV T antigens in MCC oncogenesis, identifying additional factors involved in this process, and for screening potential MCPyV+ MCC therapeutic strategies.

scholarly journals Loss of Nitric Oxide Induces Fibrogenic Response in Organotypic 3D Co-Culture of Mammary Epithelia and Fibroblasts—An Indicator for Breast Carcinogenesis

Cancers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 2815
Author(s):  
Gang Ren ◽  
Xunzhen Zheng ◽  
Vandana Sharma ◽  
Joshua Letson ◽  
Andrea L. Nestor-Kalinoski ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Culture System ◽  
Epithelial Mesenchymal Transition ◽  
Precancerous Lesions ◽  
Three Dimensional ◽  
Epidemiological Studies ◽  
Mesenchymal Transition ◽  
In Vitro Response ◽  
Mammary Epithelia

Excessive myofibroblast activation, which leads to dysregulated collagen deposition and the stiffening of the extracellular matrix (ECM), plays pivotal roles in cancer initiation and progression. Cumulative evidence attests to the cancer-causing effects of a number of fibrogenic factors found in the environment, diseases and drugs. While identifying such factors largely depends on epidemiological studies, it would be of great importance to develop a robust in vitro method to demonstrate the causal relationship between fibrosis and cancer. Here, we tested whether our recently developed organotypic three-dimensional (3D) co-culture would be suitable for that purpose. This co-culture system utilizes the discontinuous ECM to separately culture mammary epithelia and fibroblasts in the discrete matrices to model the complexity of the mammary gland. We observed that pharmaceutical deprivation of nitric oxide (NO) in 3D co-cultures induced myofibroblast differentiation of the stroma as well as the occurrence of epithelial–mesenchymal transition (EMT) of the parenchyma. Such in vitro response to NO deprivation was unique to co-cultures and closely mimicked the phenotype of NO-depleted mammary glands exhibiting stromal desmoplasia and precancerous lesions undergoing EMT. These results suggest that this novel 3D co-culture system could be utilized in the deep mechanistic studies of the linkage between fibrosis and cancer.

Effects of low doses of mifepristone on human embryo implantation process in a three-dimensional human endometrial in vitro co-culture system

Contraception ◽  
2016 ◽  
Vol 94 (2) ◽  
pp. 143-151 ◽  
Author(s):  
N.R. Boggavarapu ◽  
C. Berger ◽  
C. von Grothusen ◽  
J. Menezes ◽  
K. Gemzell-Danielsson ◽  
...  
Keyword(s):  
Human Embryo ◽  
Culture System ◽  
Embryo Implantation ◽  
Three Dimensional ◽  
Low Doses ◽  
Implantation Process

In Vitro Replication Assay for Merkel Cell Polyomavirus (MCPyV)

2015 ◽  
Vol 38 (1) ◽  
Author(s):  
Friederike Neumann ◽  
Manja Czech‐Sioli ◽  
Adam Grundhoff ◽  
Nicole Fischer
Keyword(s):  
Merkel Cell Polyomavirus ◽  
Merkel Cell ◽  
Replication Assay ◽  
In Vitro Replication

scholarly journals Physical confinement signals regulate the organization of stem cells in three dimensions

2016 ◽  
Vol 13 (123) ◽  
pp. 20160613 ◽  
Author(s):  
Sebastian V. Hadjiantoniou ◽  
David Sean ◽  
Maxime Ignacio ◽  
Michel Godin ◽  
Gary W. Slater ◽  
...  
Keyword(s):  
Stem Cells ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Cell Interactions ◽  
Three Dimensions ◽  
Cell Substrate ◽  
Cell Cell

During embryogenesis, the spherical inner cell mass (ICM) proliferates in the confined environment of a blastocyst. Embryonic stem cells (ESCs) are derived from the ICM, and mimicking embryogenesis in vitro , mouse ESCs (mESCs) are often cultured in hanging droplets. This promotes the formation of a spheroid as the cells sediment and aggregate owing to increased physical confinement and cell–cell interactions. In contrast, mESCs form two-dimensional monolayers on flat substrates and it remains unclear if the difference in organization is owing to a lack of physical confinement or increased cell–substrate versus cell–cell interactions. Employing microfabricated substrates, we demonstrate that a single geometric degree of physical confinement on a surface can also initiate spherogenesis. Experiment and computation reveal that a balance between cell–cell and cell–substrate interactions finely controls the morphology and organization of mESC aggregates. Physical confinement is thus an important regulatory cue in the three-dimensional organization and morphogenesis of developing cells.

scholarly journals Age-Specific Seroprevalences of Merkel Cell Polyomavirus, Human Polyomaviruses 6, 7, and 9, and Trichodysplasia Spinulosa-Associated Polyomavirus

2013 ◽  
Vol 20 (3) ◽  
pp. 363-368 ◽  
Author(s):  
Jérôme T. J. Nicol ◽  
Rémy Robinot ◽  
Audrey Carpentier ◽  
Giovanni Carandina ◽  
Elisa Mazzoni ◽  
...  
Keyword(s):  
Merkel Cell Polyomavirus ◽  
Human Polyomavirus ◽  
Merkel Cell ◽  
High Antibody ◽  
Specific Antibodies ◽  
Virus Like Particle ◽  
Antibody Titers ◽  
Immunosorbent Assays ◽  
Human Polyomaviruses

ABSTRACTSix new human polyomaviruses have been identified since 2008 (Merkel cell polyomavirus [MCPyV], human polyomavirus 6 [HPyV6], HPyV7, HPyV9, trichodysplasia spinulosa polyomavirus [TSPyV], and Malawi polyomavirus [MWPyV]). The presence of specific antibodies against MCPyV, HPyV6, HPyV7, HPyV9, and TSPyV in 828 Italian subjects aged 1 to 100 years was investigated by virus-like particle-based enzyme-linked immunosorbent assays (ELISAs). The findings indicate that all of these new polyomaviruses circulate widely in humans, with seroprevalences in adulthood ranging from 39.4% for HPyV9 to 87.1% for MCPyV, and that primary exposure is most intense in childhood, with the exception of HPyV7 and HPyV9, for which the seroprevalence increased throughout life. The proportion of subjects with high antibody titers was found to increase with age for MCPyV and to decrease with age for TSPyV.

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