scholarly journals Long-Term Maintenance of Viable Adult Rat Sertoli Cells Able to Establish Testis Barrier Components and Function in Response to Androgens

Cells ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2405
Author(s):  
Hassan Kabbesh ◽  
Muhammad A. Riaz ◽  
Alexandra D. Jensen ◽  
Georgios Scheiner-Bobis ◽  
Lutz Konrad
Keyword(s):  
Tight Junctions ◽  
Transforming Growth Factor Beta ◽  
Sertoli Cells ◽  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Bone Morphogenetic Protein 2 ◽  
Adult Rat ◽  
Vitro Model

A protocol for the isolation and long-term propagation of adult rat Sertoli cells (SCs) using conditional reprogramming (CR) was developed and the formation of tight junctions as an in vitro model for the blood testis barrier (BTB) was studied. Three pure primary SC lines were isolated successfully and maintained for several months without significant changes in expression levels of SC-typical markers such as SRY-box transcription factor 9 (SOX9), transferrin, clusterin, androgen receptor (AR), and GATA binding protein 1 (GATA1). In addition to AR expression, the tight junction proteins, zonula occludens-1 (ZO-1) and the junctional adhesion molecule-3 (JAM-3), were upregulated and the SC barrier integrity was enhanced by testosterone. Peritubular/myoid cells did not increase the tightness of the SC. The cytokines, interleukin-6 (IL-6), bone morphogenetic protein-2 (BMP2), and transforming growth factor beta-3 (TGF-β3), negatively affected the tightness of the SC barrier. We have established a protocol for the isolation and long-term propagation of highly pure primary adult rat SCs, which are able to respond to androgen treatments, to form tight junctions and to maintain the mRNA expression of SC-specific genes. By applying this new method, adult SCs can now be analyzed in more detail and might serve as an in vitro model for the study of many SC functions.

scholarly journals Antifibrotic effect of Ocimum basilicum L. and linalool on arecoline-induced fibrosis in human buccal fibroblasts

2018 ◽  
Vol 3 ◽  
pp. 2057178X1876447 ◽  
Author(s):  
Pooja Adtani ◽  
Narasimhan Malathi ◽  
Kannan Ranganathan ◽  
Sivaswamy Lokeswari ◽  
Alan Mathew Punnoose
Keyword(s):  
Transforming Growth Factor Beta ◽  
In Vitro Model ◽  
Transforming Growth Factor ◽  
Oral Submucous Fibrosis ◽  
Ocimum Basilicum ◽  
Morphological Alterations ◽  
Antifibrotic Effect ◽  
Submucous Fibrosis ◽  
Vitro Model

Aim: To explore Ocimum basilicum L. (sweet basil) and linalool for their antifibrotic activity in an arecoline-induced in vitro fibrotic model. Methods: Leaf extract of O. basilicum L. (LEOB) and linalool were used as experimental agents to test their antifibrogenic activity in vitro. Half-maximal inhibitory concentration (IC50) for arecoline, ethanolic LEOB, and linalool was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To evaluate the antifibrotic effect of ethanolic LEOB and linalool on pretreatment, that is, both the testing agents were added to the human buccal fibroblasts (HBFs) prior to induction with arecoline, and reverse transcriptase polymerase chain reaction (RT-PCR) was carried out to study the response of transforming growth factor beta (TGFβ), collagen 1 subtype A2 (COL1A2), and collagen 3 subtype A1 (COL3A1). To appreciate the morphological alterations in HBFs on treatment with arecoline, ethanolic LEOB, and linalool, Masson’s trichrome staining was performed. Results: Arecoline enhanced fibrotic activity by upregulating TGFβ1, COL1A2, and COL3A1 levels, whereas ethanolic LEOB and linalool on pretreatment significantly downregulated the increased levels of TGFβ1, COL1A2, and COL3A1 in primary HBF cell cultures. Conclusion and implication to clinic: Both ethanolic LEOB and linalool exhibited significant antifibrotic activity in an in vitro model. Further studies in an in vitro model can help attain a foundation for an herbal formulation in gel form that can be prescribed to patients diagnosed with oral submucous fibrosis for topical application. It can also be used synergistically with Western medicine.

scholarly journals A Pilot Study To Establish an In Vitro Model To Study Premature Intestinal Epithelium and Gut Microbiota Interactions

mSphere ◽  
2021 ◽  
Author(s):  
Justin Gibbons ◽  
Ji Youn Yoo ◽  
Tina Mutka ◽  
Maureen Groer ◽  
Thao T. B. Ho
Keyword(s):  
Cell Culture ◽  
In Vitro Model ◽  
Bacterial Flora ◽  
Mucosal Barrier ◽  
Clinical Observations ◽  
Culture Models ◽  
Vitro Model ◽  
Cell Culture Models

The gut bacterial flora influences the development of the immune system and long-term health outcomes in preterm infants. Studies of the mechanistic interactions between the gut bacteria and mucosal barrier are limited to clinical observations, animal models, and in vitro cell culture models for this vulnerable population.

scholarly journals Regulation of Decellularized Tissue Remodeling via Scaffold-Mediated Lentiviral Delivery in Anatomically-Shaped Osteochondral Constructs

2018 ◽  
Author(s):  
Christopher R. Rowland ◽  
Katherine A. Glass ◽  
Adarsh R. Ettyreddy ◽  
Catherine C. Gloss ◽  
Jared Matthews ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Protein Delivery ◽  
Spatial Organization ◽  
Transforming Growth Factor ◽  
Interleukin 1 ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Site Specific ◽  
Engineer Cartilage

AbstractCartilage-derived matrix (CDM) has emerged as a promising scaffold material for tissue engineering of cartilage and bone due to its native chondroinductive capacity and its ability to support endochondral ossification. Because it consists of native tissue, CDM can undergo cellular remodeling, which can promote integration with host tissue and enables it to be degraded and replaced by neotissue over time. However, enzymatic degradation of decellularized tissues can occur unpredictably and may not allow sufficient time for mechanically competent tissue to form, especially in the harsh inflammatory environment of a diseased joint. The goal of the current study was to engineer cartilage and bone constructs with the ability to inhibit aberrant inflammatory processes caused by the cytokine interleukin-1 (IL-1), through scaffold-mediated delivery of lentiviral particles containing a doxycycline-inducible IL-1 receptor antagonist (IL-1Ra) transgene on anatomically-shaped CDM constructs. Additionally, scaffold-mediated lentiviral gene delivery was used to facilitate spatial organization of simultaneous chondrogenic and osteogenic differentiation via site-specific transduction of a single mesenchymal stem cell (MSC) population to overexpress either chondrogenic, transforming growth factor-beta 3 (TGF-β3), or osteogenic, bone morphogenetic protein-2 (BMP-2), transgenes. Controlled induction of IL-1Ra expression protected CDM hemispheres from inflammation-mediated degradation, and supported robust bone and cartilage tissue formation even in the presence of IL-1. In the absence of inflammatory stimuli, controlled cellular remodeling was exploited as a mechanism for fusing concentric CDM hemispheres overexpressing BMP-2 and TGF-β3 into a single bi-layered osteochondral construct. Our findings demonstrate that site-specific delivery of inducible and tunable transgenes confers spatial and temporal control over both CDM scaffold remodeling and neotissue composition. Furthermore, these constructs provide a microphysiological, in vitro, joint, organoid model with site-specific, tunable, and inducible protein delivery systems for examining the spatiotemporal response to pro-anabolic and/or inflammatory signaling across the osteochondral interface.

scholarly journals New Gene Markers Involved in Molecular Processes of Tissue Repair, Response to Wounding and Regeneration Are Differently Expressed in Fibroblasts from Porcine Oral Mucosa during Long-Term Primary Culture

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1938
Author(s):  
Artur Bryja ◽  
Patrycja Sujka-Kordowska ◽  
Aneta Konwerska ◽  
Sylwia Ciesiółka ◽  
Maria Wieczorkiewicz ◽  
...  
Keyword(s):  
Wound Healing ◽  
Epithelial Cells ◽  
Expression Profile ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Primary Cultures ◽  
Gene Markers ◽  
Morphological Process

The mechanisms of wound healing and vascularization are crucial steps of the complex morphological process of tissue reconstruction. In addition to epithelial cells, fibroblasts play an important role in this process. They are characterized by dynamic proliferation and they form the stroma for epithelial cells. In this study, we have used primary cultures of oral fibroblasts, obtained from porcine buccal mucosa. Cells were maintained long-term in in vitro conditions, in order to investigate the expression profile of the molecular markers involved in wound healing and vascularization. Based on the Affymetrix assays, we have observed three ontological groups of markers as wound healing group, response to wounding group and vascularization group, represented by different genes characterized by their expression profile during long-term primary in vitro culture (IVC) of porcine oral fibroblasts. Following the analysis of gene expression in three previously identified groups of genes, we have identified that transforming growth factor beta 1 (TGFB1), ITGB3, PDPN, and ETS1 are involved in all three processes, suggesting that these genes could be recognized as markers of repair specific for oral fibroblasts within the porcine mucosal tissue.

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