scholarly journals Pseudopterosin and O-Methyltylophorinidine Suppress Cell Growth in a 3D Spheroid Co-Culture Model of Pancreatic Ductal Adenocarcinoma

2020 ◽  
Vol 7 (2) ◽  
pp. 57
Author(s):  
Bailu Xie ◽  
Jan Hänsel ◽  
Vanessa Mundorf ◽  
Janina Betz ◽  
Irene Reimche ◽  
...  
Keyword(s):  
Pancreatic Ductal Adenocarcinoma ◽  
In Vitro Model ◽  
Type I Collagen ◽  
Ductal Adenocarcinoma ◽  
Pancreatic Tumors ◽  
Type I ◽  
Culture In Vitro ◽  
Drug Candidates ◽  
Vitro Model

Current therapies for treating pancreatic ductal adenocarcinoma (PDAC) are largely ineffective, with the desmoplastic environment established within these tumors being considered a central issue. We established a 3D spheroid co-culture in vitro model using a PDAC cell line (either PANC-1 or Capan-2), combined with stellate cells freshly isolated from pancreatic tumors (PSC) or hepatic lesions (HSC), and human type I collagen to analyze the efficiency of the chemotherapeutic gemcitabine (GEM) as well as two novel drug candidates derived from natural products: pseudopterosin (PsA-D) and O-methyltylophorinidine (TYLO). Traditional 2D in vitro testing of these agents for cytotoxicity on PANC-1 demonstrated IC50 values of 4.6 (±0.47) nM, 34.02 (±1.35) µM, and 1.99 (±0.13) µM for Tylo, PsA-D, and GEM, respectively; these values were comparable for Capan-2: 5.58 (±1.74) nM, 33.94 (±1.02) µM, and 0.41 (±0.06) µM for Tylo, PsA-D, and GEM, respectively. Importantly, by assessing the extent of viable cells within 3D co-culture spheroids of PANC-1 with PSC or HSC, we could demonstrate a significant lack of efficacy for GEM, while TYLO remained active and PsA-D showed slightly reduced efficacy: GEM in PANC-1/PSC (IC50 = >100 µM) or PANC-1/HSC (IC50 = >100 µM) spheroids, TYLO in PANC-1/PSC (IC50 = 3.57 ± 1.30 nM) or PANC-1/HSC (IC50 = 6.39 ± 2.28 nM) spheroids, and to PsA-D in PANC-1/PSC (IC50 = 54.42 ± 12.79 µM) or PANC-1/HSC (IC50 = 51.75 ± 0.60 µM). Microscopic 3D rendering supported these cytotoxicity outcomes, showing little or no morphological spheroid structure change during this period of rapid cell death. Our results support the use of this 3D spheroid co-culture in vitro model having a desmoplastic microenvironment for the identification of possible novel chemotherapeutic drug candidates for PDAC, such as TYLO and PsA-D.

scholarly journals Anti-Photoaging Effect of Plant Extract Fermented with Lactobacillus buchneri on CCD-986sk Fibroblasts and HaCaT Keratinocytes

2020 ◽  
Vol 11 (1) ◽  
pp. 3 ◽  
Author(s):  
Yun-Mi Kang ◽  
Chul-Hee Hong ◽  
Sa-Haeng Kang ◽  
Dong-Seok Seo ◽  
Seong-Oh Kim ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Type I Collagen ◽  
Dermal Fibroblasts ◽  
Epidermal Keratinocytes ◽  
Type I ◽  
Mrna Levels ◽  
Protective Effects ◽  
Lactobacillus Buchneri ◽  
Vitro Model

Ultraviolet (UV) exposure triggers the abnormal production of reactive oxygen (ROS) species and the expression of matrix metalloproteinases (MMPs) that are responsible for photoaging. Probiotics are widely used in healthcare and for immune enhancement. One probiotic, Lactobacillus buchneri is found in Kimchi. This study was aimed at assessing the anti-photoaging effect of plant extracts fermented with L. buchneri (PELB) to develop functional cosmetics. We investigated the anti-photoaging effect of PELB in a UVB-induced photoaging in vitro model and selected effective extracts using the elastase inhibition assay, ELISA for Type I procollagen and collagenase-1, and quantitative real time PCR. Normal human dermal fibroblasts and epidermal keratinocytes were pre-treated with PELB and exposed to UVB. We found that PELB decreased elastase activity and increased type I collagen expression in a UVB-induced photoaging in vitro model. In addition, PELB greatly reduced collagenase activity and MMP mRNA levels in a UVB-induced photoaging in vitro model. Furthermore, PELB promoted the expression of moisture factor and anti-oxidant enzymes in a UVB-induced photoaging in vitro model. These results indicated that the PELB could be potential candidates for the protective effects against UVB-induced photoaging. Overall, these results suggest that PELB might be useful natural components of cosmetic products.

A serum-free in vitro model of renal microvessel development

1998 ◽  
Vol 274 (6) ◽  
pp. F1150-F1160 ◽  
Author(s):  
Lisa M. Antes ◽  
Monica M. Villar ◽  
Sylvia Decker ◽  
Roberto F. Nicosia ◽  
Dean A. Kujubu
Keyword(s):  
Growth Factors ◽  
Endothelial Cell ◽  
In Vitro Model ◽  
Type I Collagen ◽  
Type I ◽  
Renal Vasculature ◽  
Serum Free ◽  
Polypeptide Growth Factors ◽  
Vitro Model

The differentiation and organization of the embryonic renal vasculature is a crucial event in renal development. To study this process, we developed a serum-free in vitro model of renal microvessel development. Mouse embryonic kidney explants, when embedded specifically in type I collagen, demonstrate outgrowth of microvascular structures when stimulated by the phorbol ester 12- O-tetradecanoylphorbol 13-acetate (TPA, 10–50 ng/ml). Other polypeptide growth factors stimulated little, if any, microvessel outgrowth from the explants. Similar outgrowths were not observed when other embryonic tissue explants were used. The number of microvessels observed depended on the gestational age of the explants. We hypothesize that TPA induces the in situ differentiation of metanephric mesenchymal cells into endothelial cell precursors and that specific matrix proteins and cell-matrix interactions are necessary for the organization of these precursors into microvessels. Our model will allow us to examine in detail the responsiveness of metanephric kidney cells to both growth factors and extracellular matrix molecules and to understand how they influence renal endothelial cell differentiation.

scholarly journals In Vitro Model of Bartonella henselae-Induced Angiogenesis

2004 ◽  
Vol 72 (12) ◽  
pp. 7315-7317 ◽  
Author(s):  
James E. Kirby
Keyword(s):  
In Vitro Model ◽  
Type I Collagen ◽  
Bartonella Henselae ◽  
Collagen Matrix ◽  
In Vitro Models ◽  
Type I ◽  
Blood Vessel Formation ◽  
Vitro Model

ABSTRACT Bartonella henselae is a gram-negative pathogen that causes angiogenesis. Here, I establish in vitro models to study Bartonella-induced blood vessel formation. I found that B. henselae induces long-term endothelial survival and tubular differentiation within type I collagen matrix.

scholarly journals Evaluation In Vivo and In Vitro of the Antioxidant, Antinociceptive, and Anti-Inflammatory Activities of Biflavonoids From Ouratea hexasperma and O. ferruginea

2019 ◽  
Vol 14 (6) ◽  
pp. 1934578X1985680 ◽  
Author(s):  
Poliana de Araujo Oliveira ◽  
Queli Cristina Fidelis ◽  
Thayane Ferreira da Costa Fernandes ◽  
Milene Conceição de Souza ◽  
Dayane Magalhães Coutinho ◽  
...  
Keyword(s):  
Type I Collagen ◽  
In Vivo Model ◽  
Type I ◽  
Anti Inflammatory ◽  
Vitro Model ◽  
Factor Α ◽  
Necrosis Factor

Ouratea species are used for the treatment of inflammation-related diseases such as rheumatism and arthritic disorders. The Ouratea genus is a rich source of flavonoids and bioflavonoids and for this reason we evaluated the effects of the biflavonoid fractions from the leaves of O. hexasperma (OHME) and O. ferruginea (OFME) in the in vivo model of complete Freund’s adjuvant (CFA)-induced arthritis and in the in vitro model of oxidative stress and cellular viability. The CFA-induced arthritis model in rats was followed by paw volume, articular incapacitation and Randall-selitto models, as well as quantification of cytokines and serum C-terminal telopeptide of type I collagen levels. OHME and OFME demonstrated antinociceptive and anti-inflammatory activities, as well as improvement in articular incapacity and reduction in levels of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α, and type 1 collagen, and increased cell viability. No adverse effects were observed. The results suggest that OHME and OFME can reduce inflammation and bone resorption besides their antioxidant action.

scholarly journals Development and Validation of Fluorescently Labeled, Functional Type I Collagen Molecules

2021 ◽  
Author(s):  
Seyed Mohammad Siadat ◽  
Monica E Susilo ◽  
Jeffrey A Paten ◽  
Alexandra A Silverman ◽  
Charles A DiMarzio ◽  
...  
Keyword(s):  
Type I Collagen ◽  
De Novo ◽  
Type I ◽  
Growth And Remodeling ◽  
Fibril Structure ◽  
Collagen Molecules ◽  
Vitro Model ◽  
Development And Validation ◽  
Network Morphology

While de novo collagen fibril formation is well-studied, there are few investigations into the growth and remodeling of extant fibrils, where molecular collagen incorporation into and erosion from the fibril surface must delicately balance during fibril growth and remodeling. Observing molecule/fibril interactions is difficult, requiring the tracking of molecular dynamics while, at the same time, minimizing the effect of the observation on fibril structure and assembly. To address the observation-interference problem, exogenous collagen molecules are tagged with small fluorophores and the fibrillogenesis kinetics of labeled collagen molecules as well as the structure and network morphology of assembled fibrils are quantified for the first time. While excessive labeling significantly disturbs fibrillogenesis kinetics and network morphology of assembled fibrils, adding less than ~1.2 labels preserves them. Applications of the functional, labeled collagen probe are demonstrated in both cellular and acellular systems. The functional, labelled collagen associates strongly with native fibrils and when added to an in vitro model of corneal stromal development, is endocytosed rapidly by cells and is translocated into synthesized matrix networks within 24 hours.

A Simple In Vitro Model of Mechanical Injury of Confluent Cultured Endothelial Cells to Study Quantitatively the Repair Process

1986 ◽  
Vol 56 (02) ◽  
pp. 232-235 ◽  
Author(s):  
Claude Klein-Soyer ◽  
Alain Beretz ◽  
Régine Millon-Collard ◽  
Joseph Abecassis ◽  
Jean-Pierre Cazenave
Keyword(s):  
Endothelial Cells ◽  
In Vitro Model ◽  
Repair Process ◽  
Mechanical Injury ◽  
Brain Extract ◽  
Type I ◽  
Culture Dish ◽  
Vitro Model ◽  
Adhesive Proteins

SummaryA model of in vitro mechanical injury of confluent human endothelial cells (EC) in culture was developed. Human EC were obtained from umbilical veins and grown to confluence. Application on the EC monolayer of a calibrated disk of cellulose poly acetate paper resulted in removal of the EC, leaving a continuous subendothelial extracellular matrix (ECM) on the culture dish. The regeneration time depended on the original size of the lesion. Regeneration was similar with EC grown on different substrates such as human fibronectin, human subendothelial ECM, bovine collagen type I or surfaces coated with Transglutine®, a surgical glue containing adhesive proteins. A human brain extract containing growth factor activity accelerated significantly the repair of the lesion, especially at low serum concentration. This simple in vitro model of mechanical injury allows the quantitative study of the effects of matrices, growth factors and pharmacological agents on the repair process.

scholarly journals Novel in vitro model of urea cycle disorder using citrullinemia type I patient-derived iPSC s

HPB ◽  
2016 ◽  
Vol 18 ◽  
pp. e300
Author(s):  
E.Y. Yoshitoshi ◽  
T. Toyoda ◽  
K. Yasuda ◽  
M. Kotaka ◽  
K. Okita ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Urea Cycle ◽  
Urea Cycle Disorder ◽  
Type I ◽  
Vitro Model ◽  
Citrullinemia Type I
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