Establishment of in vitro cellular model predicting histocompatibity in allograft

2001 ◽  
Vol 21 (4) ◽  
pp. 277-279
Author(s):  
Hao Youhua ◽  
Wu Xiongwen ◽  
Liang Zhihui ◽  
Xiong Ping ◽  
Jiang Xiaodan ◽  
...  
Keyword(s):  
Cellular Model

scholarly journals A rapid and robust method for the cryopreservation of human granulosa cells

2021 ◽  
Author(s):  
Sarah Beschta ◽  
Katja Eubler ◽  
Nancy Bohne ◽  
Ignasi Forne ◽  
Dieter Berg ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Large Scale ◽  
Ovarian Function ◽  
Oocyte Retrieval ◽  
Cell Contact ◽  
Cellular Model ◽  
Preovulatory Follicle ◽  
Human Granulosa Cells ◽  
Ovarian Cells

AbstractHuman primary granulosa cells (GCs) derived from women undergoing oocyte retrieval can be cultured and used as a cellular model for the study of human ovarian function. In vitro, they change rapidly, initially resembling cells of the preovulatory follicle and then cells of the corpus luteum. They are derived from individual patients, whose different medical history, lifestyle and age lead to heterogeneity. Thus, cells can rarely be ideally matched for cellular experiments or, if available, only in small quantities. We reasoned that cryopreservation of human GCs may be helpful to improve this situation. Previous studies indicated the feasibility of such an approach, but low survival of human GCs was reported, and effects on human GC functionality were only partially evaluated. We tested a slow freezing protocol (employing FCS and DMSO) for human GCs upon isolation from follicular fluid. We compared cryopreserved and subsequently thawed cells with fresh, non-cryopreserved cells from the same patients. About 80% of human GCs survived freezing/thawing. No differences were found in cell morphology, survival rate in culture, or transcript levels of mitochondrial (COX4, OPA1, TOMM20), steroidogenic (CYP11A1, CYP19A1) or cell–cell contact genes (GJA1) between the two groups in cells cultured for 1–5 days. A proteomic analysis revealed no statistically significant change in the abundance of a total of 5962 proteins. The two groups produced comparable basal levels of progesterone and responded similarly to hCG with elevation of progesterone. Taken together, our results show this to be a rapid and readily available method for the cryopreservation of human GCs. We anticipate that it will allow future large-scale experiments and may thereby improve cellular studies with human ovarian cells.

scholarly journals A translational cellular model for the study of peritubular cells of the testis

Reproduction ◽  
2020 ◽  
Vol 160 (2) ◽  
pp. 259-268 ◽  
Author(s):  
Nina Schmid ◽  
Annika Missel ◽  
Stoyan Petkov ◽  
Jan B Stöckl ◽  
Florian Flenkenthaler ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Replicative Senescence ◽  
Smooth Muscle Actin ◽  
Proteome Analysis ◽  
Inflammatory Factors ◽  
Seminiferous Tubules ◽  
Cellular Model ◽  
Mechanistic Studies ◽  
Peritubular Cells

Testicular peritubular cells (TPCs) are smooth muscle-like cells, which form a compartment surrounding the seminiferous tubules. Previous studies employing isolated human testicular peritubular cells (HTPCs) indicated that their roles in the testis go beyond sperm transport and include paracrine and immunological contributions. Peritubular cells from a non-human primate (MKTPCs), the common marmoset monkey, Callithrix jacchus, share a high degree of homology with HTPCs. However, like their human counterparts these cells age in vitro and replicative senescence limits in-depth functional or mechanistic studies. Therefore, a stable cellular model was established. MKTPCs of a young adult animal were immortalized by piggyBac transposition of human telomerase (hTERT), that is, without the expression of viral oncogenes. Immortalized MKTPCs (iMKTPCs) grew without discernable changes for more than 50 passages. An initial characterization revealed typical genes expressed by peritubular cells (androgen receptor (AR), smooth-muscle actin (ACTA2), calponin (CNN1)). A proteome analysis of the primary MKTPCs and the derived immortalized cell line confirmed that the cells almost completely retained their phenotype. To test whether they respond in a similar way as HTPCs, iMKTPCs were challenged with forskolin (FSK) and ATP. As HTPCs, they showed increased expression level of the StAR protein (StAR) after FSK stimulation, indicating steroidogenic capacity. ATP increased the expression of pro-inflammatory factors (e.g. IL1B; CCL7), as it is the case in HTPCs. Finally, we confirmed that iMKTPCs can efficiently be transfected. Therefore, they represent a highly relevant translational model, which allows mechanistic studies for further exploration of the roles of testicular peritubular cells.

scholarly journals An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment

2020 ◽  
Vol 9 (7) ◽  
pp. 32
Author(s):  
Jingwen Cai ◽  
Kristin Perkumas ◽  
W. Daniel Stamer ◽  
Yutao Liu
Keyword(s):  
Endothelial Cells ◽  
Cellular Model ◽  
Schlemm’S Canal ◽  
Schlemm's Canal

Insight into antioxidant properties of milk‐derived bioactive peptides in vitro and in a cellular model

2019 ◽  
Vol 25 (5) ◽  
pp. e3162 ◽  
Author(s):  
Federica Tonolo ◽  
Laura Moretto ◽  
Stefania Ferro ◽  
Alessandra Folda ◽  
Valeria Scalcon ◽  
...  
Keyword(s):  
Bioactive Peptides ◽  
Antioxidant Properties ◽  
Cellular Model ◽  
Insight Into

scholarly journals Toward a 3D Cellular Model for Studying In Vitro the Outcome of Photodynamic Treatments: Accounting for the Effects of Tissue Complexity

2013 ◽  
Vol 19 (15-16) ◽  
pp. 1665-1674 ◽  
Author(s):  
Mireia Alemany-Ribes ◽  
María García-Díaz ◽  
Marta Busom ◽  
Santi Nonell ◽  
Carlos E. Semino
Keyword(s):  
Cellular Model

scholarly journals A Novel Cellular Model to Study Angiotensin II AT2 Receptor Function in Breast Cancer Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Sylvie Rodrigues-Ferreira ◽  
Marina Morel ◽  
Rosana I. Reis ◽  
Françoise Cormier ◽  
Véronique Baud ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Fluorescent Protein ◽  
Receptor Activation ◽  
At1 Receptor ◽  
At2 Receptor ◽  
Cellular Model ◽  
Selective Antagonist

Recent studies have highlighted the AT1 receptor as a potential therapeutic target in breast cancer, while the role of the AT2 subtype in this disease has remained largely neglected. The present study describes the generation and characterization of a new cellular model of human invasive breast cancer cells (D3H2LN-AT2) stably expressing high levels of Flag-tagged human AT2 receptor (Flag-hAT2). These cells exhibit high-affinity binding sites for AngII, and total binding can be displaced by the AT2-selective antagonist PD123319 but not by the AT1-selective antagonist losartan. Of interest, high levels of expression of luciferase and green fluorescent protein make these cells suitable for bioluminescence and fluorescence studies in vitro and in vivo. We provide here a novel tool to investigate the AT2 receptor functions in breast cancer cells, independently of AT1 receptor activation.

scholarly journals The genotype 3-specific hepatitis C virus core protein residue phenylalanine 164 increases steatosis in an in vitro cellular model

Gut ◽  
2007 ◽  
Vol 56 (9) ◽  
pp. 1302-1308 ◽  
Author(s):  
C Hourioux ◽  
R Patient ◽  
A Morin ◽  
E Blanchard ◽  
A Moreau ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Core Protein ◽  
Cellular Model ◽  
Genotype 3 ◽  
Virus Core

scholarly journals Human Mesenchymal Stem Cell Combined with a New Strontium-Enriched Bioactive Glass: An ex-vivo Model for Bone Regeneration

Materials ◽  
2019 ◽  
Vol 12 (21) ◽  
pp. 3633 ◽  
Author(s):  
Devis Bellucci ◽  
Elena Veronesi ◽  
Valentina Strusi ◽  
Tiziana Petrachi ◽  
Alba Murgia ◽  
...  
Keyword(s):  
Bioactive Glass ◽  
Ex Vivo ◽  
Human Mesenchymal Stem Cell ◽  
Cellular Model ◽  
Ex Vivo Model ◽  
Bone Differentiation ◽  
Biological Potential ◽  
Orthopedic Applications ◽  
First Time

A 3D cellular model that mimics the potential clinical application of a biomaterial is here applied for the first time to a bioactive glass, in order to assess its biological potential. A recently developed bioactive glass (BGMS10), whose composition contained strontium and magnesium, was produced in the form of granules and fully investigated in terms of biocompatibility in vitro. Apart from standard biological characterization (Simulated Body Fluid (SBF) testing and biocompatibility as per ISO10993), human bone marrow mesenchymal stromal/stem cells (BM-MSCs) were used to investigate the performance of the bioactive glass granules in an innovative 3D cellular model. The results showed that BGMS10 supported human BM-MSCs adhesion, colonization, and bone differentiation. Thus, bioactive glass granules seem to drive osteogenic differentiation and thus look particularly promising for orthopedic applications, bone tissue engineering and regenerative medicine.

scholarly journals In vitro comparison of hydroxocobalamin (B12a) and the mitochondrial directed therapy by a succinate prodrug in a cellular model of cyanide poisoning

2020 ◽  
Vol 7 ◽  
pp. 1263-1271 ◽  
Author(s):  
Shawn Owiredu ◽  
Abhay Ranganathan ◽  
John C. Greenwood ◽  
Sarah Piel ◽  
Joanna I. Janowska ◽  
...  
Keyword(s):  
Cellular Model ◽  
Cyanide Poisoning ◽  
In Vitro Comparison

scholarly journals TET-catalyzed 5-carboxylcytosine promotes CTCF binding to suboptimal sequences genome-wide

2018 ◽  
Author(s):  
Kyster K. Nanan ◽  
David M. Sturgill ◽  
Maria F. Prigge ◽  
Morgan Thenoz ◽  
Allissa A. Dillman ◽  
...  
Keyword(s):  
Binding Sites ◽  
Genomic Dna ◽  
Multiple Roles ◽  
Ctcf Binding ◽  
Cellular Model ◽  
Potential Mechanism ◽  
Dynamic Regulation ◽  
Genome Wide

SummaryThe mechanisms supporting dynamic regulation of CTCF binding sites remain poorly understood. Here we describe the TET-catalyzed 5-methylcytosine derivative, 5-carboxylcytosine (5caC) as a factor driving new CTCF binding within genomic DNA. Through a combination of in vivo and in vitro approaches, we reveal that 5caC generally strengthens CTCF association with DNA and facilitates binding to suboptimal sequences. Dramatically, profiling of CTCF binding in a cellular model that accumulates genomic 5caC identified ∼13,000 new CTCF sites. The new sites were enriched for overlapping 5caC and were marked by an overall reduction in CTCF motif strength. As CTCF has multiple roles in gene expression, these findings have wide-reaching implications and point to induced 5caC as a potential mechanism to achieve differential CTCF binding in cells.

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