An in vitro screening method for probiotics with antidepressant-like effect using the enterochromaffin cell model

2021 ◽  
Author(s):  
Peijun Tian ◽  
Huiyue Zhu ◽  
Renying Zou ◽  
Qinming Kong ◽  
Mengshu Xu ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Cell Model ◽  
Screening Method ◽  
In Vitro Screening ◽  
Enterochromaffin Cell

Screening the probiotics with antidepressant-like effects through measuring the bacteria stimulated Tph1 mRNA expression and 5-HTP/5-HT secretion in the enterochromaffin cell model RIN14B.

Development of an in vitro screening method for safety evaluation of nanomaterials

2009 ◽  
Vol 19 (1) ◽  
pp. 19-27 ◽  
Author(s):  
Atsuko Matsuoka ◽  
Agneta Önfelt ◽  
Yoshie Matsuda ◽  
Ryusuke Nakaoka ◽  
Yuji Haishima ◽  
...  
Keyword(s):  
Screening Method ◽  
Safety Evaluation ◽  
In Vitro Screening

scholarly journals Generation of a hTERT-Immortalized Human Sertoli Cell Model to Study Transporter Dynamics at the Blood-Testis Barrier

Pharmaceutics ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1005
Author(s):  
Raymond K. Hau ◽  
Siennah R. Miller ◽  
Stephen H. Wright ◽  
Nathan J. Cherrington
Keyword(s):  
Mrna Expression ◽  
Cell Model ◽  
In Vitro Models ◽  
Replication Capacity ◽  
Seminiferous Tubules ◽  
Difference Threshold ◽  
Transport Pathways ◽  
Equilibrative Nucleoside Transporters ◽  
Blood Testis Barrier

The blood-testis barrier (BTB) formed by adjacent Sertoli cells (SCs) limits the entry of many chemicals into seminiferous tubules. Differences in rodent and human substrate-transporter selectivity or kinetics can misrepresent conclusions drawn using rodent in vitro models. Therefore, human in vitro models are preferable when studying transporter dynamics at the BTB. This study describes a hTERT-immortalized human SC line (hT-SerC) with significantly increased replication capacity and minor phenotypic alterations compared to primary human SCs. Notably, hT-SerCs retained similar morphology and minimal changes to mRNA expression of several common SC genes, including AR and FSHR. The mRNA expression of most xenobiotic transporters was within the 2-fold difference threshold in RT-qPCR analysis with some exceptions (OAT3, OCT3, OCTN1, OATP3A1, OATP4A1, ENT1, and ENT2). Functional analysis of the equilibrative nucleoside transporters (ENTs) revealed that primary human SCs and hT-SerCs predominantly express ENT1 with minimal ENT2 expression at the plasma membrane. ENT1-mediated uptake of [3H] uridine was linear over 10 min and inhibited by NBMPR with an IC50 value of 1.35 ± 0.37 nM. These results demonstrate that hT-SerCs can functionally model elements of transport across the human BTB, potentially leading to identification of other transport pathways for xenobiotics, and will guide drug discovery efforts in developing effective BTB-permeable compounds.

AN IN-VITRO SCREENING METHOD TO STUDY THE ACTIVITY POTENTIAL OF BIOFERTILIZERS BASED ONTRICHODERMAANDBACILLUSSP.

2013 ◽  
Vol 36 (9) ◽  
pp. 1439-1452 ◽  
Author(s):  
Zafrin Akter ◽  
Markus Weinmann ◽  
Günter Neumann ◽  
Volker Römheld
Keyword(s):  
Screening Method ◽  
In Vitro Screening

scholarly journals Curcumin Alleviates the Senescence of Canine Bone Marrow Mesenchymal Stem Cells during In Vitro Expansion by Activating the Autophagy Pathway

2021 ◽  
Vol 22 (21) ◽  
pp. 11356
Author(s):  
Jiaqiang Deng ◽  
Ping Ouyang ◽  
Weiyao Li ◽  
Lijun Zhong ◽  
Congwei Gu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Mrna Expression ◽  
Light Chain ◽  
Therapeutic Potential ◽  
Cell Model ◽  
Biological Aging ◽  
Aging Effects

Senescence in mesenchymal stem cells (MSCs) not only hinders the application of MSCs in regenerative medicine but is also closely correlated with biological aging and the development of degenerative diseases. In this study, we investigated the anti-aging effects of curcumin (Cur) on canine bone marrow-derived MSCs (cBMSCs), and further elucidated the potential mechanism of action based on the modulation of autophagy. cBMSCs were expanded in vitro with standard procedures to construct a cell model of premature senescence. Our evidence indicates that compared with the third passage of cBMSCs, many typical senescence-associated phenotypes were observed in the sixth passage of cBMSCs. Cur treatment can improve cBMSC survival and retard cBMSC senescence according to observations that Cur (1 μM) treatment can improve the colony-forming unit-fibroblasts (CFU-Fs) efficiency and upregulated the mRNA expression of pluripotent transcription factors (SOX-2 and Nanog), as well as inhibiting the senescence-associated beta-galactosidase (SA-β-gal) activities and mRNA expression of the senescence-related markers (p16 and p21) and pro-inflammatory molecules (tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)). Furthermore, Cur (0.1 μM~10 μM) was observed to increase autophagic activity, as identified by upregulation of microtubule-associated protein 1 light chain 3 (LC3), unc51-like autophagy-activating kinase-1 (ULK1), autophagy-related gene (Atg) 7 and Atg12, and the generation of type II of light chain 3 (LC3-II), thereby increasing autophagic vacuoles and acidic vesicular organelles, as well as causing a significant decrease in the p62 protein level. Moreover, the autophagy activator rapamycin (RAP) and Cur were found to partially ameliorate the senescent features of cBMSCs, while the autophagy inhibitor 3-methyladenine (3-MA) was shown to aggravate cBMSCs senescence and Cur treatment was able to restore the suppressed autophagy and counteract 3-MA-induced cBMSC senescence. Hence, our study highlights the important role of Cur-induced autophagy and its effects for ameliorating cBMSC senescence and provides new insight for delaying senescence and improving the therapeutic potential of MSCs.

scholarly journals IVIVC Assessment of Two Mouse Brain Endothelial Cell Models for Drug Screening

Pharmaceutics ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 587 ◽  
Author(s):  
Ina Puscas ◽  
Florian Bernard-Patrzynski ◽  
Martin Jutras ◽  
Marc-André Lécuyer ◽  
Lyne Bourbonnière ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Mrna Expression ◽  
Mouse Brain ◽  
Drug Screening ◽  
Cell Model ◽  
Drug Permeability ◽  
Mouse Brain Endothelial Cells ◽  
Primary Model

Since most preclinical drug permeability assays across the blood-brain barrier (BBB) are still evaluated in rodents, we compared an in vitro mouse primary endothelial cell model to the mouse b.End3 and the acellular parallel artificial membrane permeability assay (PAMPA) models for drug screening purposes. The mRNA expression of key feature membrane proteins of primary and bEnd.3 mouse brain endothelial cells were compared. Transwell® monolayer models were further characterized in terms of tightness and integrity. The in vitro in vivo correlation (IVIVC) was obtained by the correlation of the in vitro permeability data with log BB values obtained in mice for seven drugs. The mouse primary model showed higher monolayer integrity and levels of mRNA expression of BBB tight junction (TJ) proteins and membrane transporters (MBRT), especially for the efflux transporter Pgp. The IVIVC and drug ranking underlined the superiority of the primary model (r2 = 0.765) when compared to the PAMPA-BBB (r2 = 0.391) and bEnd.3 cell line (r2 = 0.019) models. The primary monolayer mouse model came out as a simple and reliable candidate for the prediction of drug permeability across the BBB. This model encompasses a rapid set-up, a fair reproduction of BBB tissue characteristics, and an accurate drug screening.

scholarly journals Simple rapid in vitro screening method for SARS-CoV-2 anti-virals that identifies potential cytomorbidity-associated false positives

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Kexin Yan ◽  
Daniel J. Rawle ◽  
Thuy T. Le ◽  
Andreas Suhrbier
Keyword(s):  
Cell Density ◽  
Drug Screening ◽  
Screening Method ◽  
False Positives ◽  
In Vitro Screening ◽  
Viral Activity ◽  
Drug Concentrations ◽  
In Vitro Drug Screening ◽  
Growth Assay

Abstract Background The international SARS-CoV-2 pandemic has resulted in an urgent need to identify new anti-viral drugs for treatment of COVID-19. The initial step to identifying potential candidates usually involves in vitro screening that includes standard cytotoxicity controls. Under-appreciated is that viable, but stressed or otherwise compromised cells, can also have a reduced capacity to replicate virus. A refinement proposed herein for in vitro drug screening thus includes a simple growth assay to identify drug concentrations that cause cellular stress or “cytomorbidity”, as distinct from cytotoxicity or loss of viability. Methods A simple rapid bioassay is presented for antiviral drug screening using Vero E6 cells and inhibition of SARS-CoV-2 induced cytopathic effects (CPE) measured using crystal violet staining. We use high cell density for cytotoxicity assays, and low cell density for cytomorbidity assays. Results The assay clearly illustrated the anti-viral activity of remdesivir, a drug known to inhibit SARS-CoV-2 replication. In contrast, nitazoxanide, oleuropein, cyclosporine A and ribavirin all showed no ability to inhibit SARS-CoV-2 CPE. Hydroxychloroquine, cyclohexamide, didemnin B, γ-mangostin and linoleic acid were all able to inhibit viral CPE at concentrations that did not induce cytotoxicity. However, these drugs inhibited CPE at concentrations that induced cytomorbidity, indicating non-specific anti-viral activity. Conclusions We describe the methodology for a simple in vitro drug screening assay that identifies potential anti-viral drugs via their ability to inhibit SARS-CoV-2-induced CPE. The additional growth assay illustrated how several drugs display anti-viral activity at concentrations that induce cytomorbidity. For instance, hydroxychloroquine showed anti-viral activity at concentrations that slow cell growth, arguing that its purported in vitro anti-viral activity arises from non-specific impairment of cellular activities. The cytomorbidity assay can therefore rapidly exclude potential false positives.

scholarly journals Efficient In Vitro Screening for Higher Soil pH Adaptability of Intersectional Hybrids in Blueberry

HortScience ◽  
2014 ◽  
Vol 49 (2) ◽  
pp. 141-144 ◽  
Author(s):  
Hirotoshi Tsuda ◽  
Hisato Kunitake ◽  
Yo Aoki ◽  
Akiko Oyama ◽  
Takuya Tetsumura ◽  
...  
Keyword(s):  
Screening Method ◽  
Multiple Shoots ◽  
Vaccinium Corymbosum ◽  
Apparent Difference ◽  
In Vitro Screening ◽  
Ph Tolerance ◽  
Short Time ◽  
Hybrid Clones ◽  
Selection Of

We tested efficient in vitro methods for screening the genotypes with higher pH tolerance using multiple shoots of intersectional hybrids between Vaccinium corymbosum ‘Spartan’ and V. bracteatum. The response of the four hybrid clones tested to different pH levels was clone-dependent in vitro. An apparent difference was found in the rooting rate among the hybrid clones even at higher pH levels; the rooting rates of JM4 (91%) at pH 8.0 indicated a significantly high value compared with other clones (JM1: 24%, JM2: 9%, JM3: 8%, ‘Spartan’: 0%). Furthermore, JM4 showed constantly high rooting rates (91% to 100%) at all pH levels with no significant differences. Similar differences in the root characters of the hybrids were also confirmed by checking the viability of roots using fluorescein diacetate (FDA)/propidium iodide (PI) staining after dipping the roots of in vitro-produced shoots in liquid medium at different pH levels for 6 hours. These results suggest that an in vitro screening method using the rooting rate of multiple shoots and the viability test of roots by FDA/PI staining as a marker could become a very useful tool for the selection of germplasm with tolerance to higher pH within a short time using small planting spaces. In addition, JM4, which showed a high rooting rate at pH 8.0, could be useful in breeding new cultivars with higher pH tolerance.

In vitro screening method for gastrotoxicity using mucin layer

2010 ◽  
Vol 196 ◽  
pp. S141
Author(s):  
J. Ha ◽  
S.C. Park ◽  
H. Kwon
Keyword(s):  
Screening Method ◽  
In Vitro Screening ◽  
Mucin Layer
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