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fibroblast coculture

Vasiliki Liakouli, Paola Cipriani, Paola Di Benedetto, Noemi Panzera, Piero Ruscitti, Ilenia Pantano, Onorina Berardicurti, Francesco Carubbi, Filomena Esteves, Georgia Mavria, Francesco Del Galdo, Roberto Giacomelli
Systemic sclerosis (SSc) is characterized by microangiopathy with impaired reparative angiogenesis and fibrosis. Epidermal Growth Factor Like-domain 7 (EGFL7), firstly described in endothelial cells plays a pivotal role in angiogenesis. Fibroblasts (FBs) are involved in vascular remodeling, under physiological and pathological conditions. In this study, we investigated: (i) the expression of EGFL7 and its miR-126 in patients affected by diffuse cutaneous SSc (dcSSc); (ii) the ability of Transforming Growth Factor-beta (TGF-β) to modulate EGFL7 expression; (iii) the ability of EGFL7 to modulate COL1A1 expression and proliferation/migration, and (iv) the functional role of EGFL7 on angiogenesis...
March 14, 2019: Scientific Reports
M N Segunda, J Bahamonde, I Muñoz, S Sepulveda, J Cortez, M De Los Reyes, J Palomino, C G Torres, O A Peralta
In vitro gamete derivation based on differentiation of germ cells (GC) from stem cells has emerged as a potential new strategy for the treatment of male infertility. This technology also has potential applications in animal reproduction as an alternative method for dissemination of elite animal genetics, production of transgenic animals, and conservation of endangered species. Mesenchymal stem cells (MSC) are multipotent progenitor cells defined by their ability to differentiate into mesodermal lineages. Under the effect of selected bioactive factors, MSC upregulate expression of pluripotent and GC specific-markers revealing their potential for GC differentiation...
February 28, 2019: Theriogenology
Asma Begum, Ross H McMillan, Yu-Tai Chang, Vesselin R Penchev, N V Rajeshkumar, Anirban Maitra, Michael G Goggins, James R Eshelman, Christopher L Wolfgang, Zeshaan A Rasheed, William Matsui
OBJECTIVE: Cancer-associated fibroblasts (CAFs) play an important role in the progression of pancreatic ductal adenocarcinoma (PDAC) by promoting tumor cell migration and drug resistance. We determined the impact of CAFs on PDAC cancer stem cells (CSCs). METHODS: Fibroblast cell lines from patients' tumors were cocultured with PDAC cells and examined for clonogenic growth and self-renewal using colony-forming assays and migration in vitro. Changes in the frequency of CSCs was determined by flow cytometry...
February 8, 2019: Pancreas
Shohei Ishikawa, Kazutoshi Iijima, Kohei Sasaki, Masaaki Kawabe, Hidenori Otsuka
In order to realize organ-on-a-chip as an effective tool for regenerative medicine and drug development, tissue-mimic cell culture methods which promote liver-specific function for long period have been developed. We have previously demonstrated that coculture of hepatocyte spheroids on fibroblasts using micropatterned substrate improved the hepatic functions due to the heterotypic cell-cell interactions and paracrine signaling from each other. In addition, hepatocyte function cultured as monolayer was also promoted in separately coculture with fibroblasts cultured as monolayer, and it is more improved in separately coculture with fibroblasts in 3D silica nonwoven fabrics...
June 1, 2019: Journal of Nanoscience and Nanotechnology
Jan B Boscheinen, Sabrina Thomann, David M Knipe, Neal DeLuca, Beatrice Schuler-Thurner, Stefanie Gross, Jan Dörrie, Niels Schaft, Christian Bach, Anette Rohrhofer, Melanie Werner-Klein, Barbara Schmidt, Philipp Schuster
Robust anti-tumor immunity requires innate as well as adaptive immune responses. We have shown that plasmacytoid dendritic cells develop killer cell-like activity in melanoma cell cocultures after exposure to the infectious but replication-deficient herpes simplex virus 1 (HSV-1) d 106S. To combine this innate effect with an enhanced adaptive immune response, the gene encoding human MelanA/MART-1 was inserted into HSV-1 d 106S via homologous recombination to increase direct expression of this tumor antigen...
2019: Frontiers in Immunology
Fernanda G Kugeratski, Samuel J Atkinson, Lisa J Neilson, Sergio Lilla, John R P Knight, Jens Serneels, Amelie Juin, Shehab Ismail, David M Bryant, Elke K Markert, Laura M Machesky, Massimiliano Mazzone, Owen J Sansom, Sara Zanivan
Intratumoral hypoxia causes the formation of dysfunctional blood vessels, which contribute to tumor metastasis and reduce the efficacy of therapeutic treatments. Blood vessels are embedded in the tumor stroma of which cancer-associated fibroblasts (CAFs) constitute a prominent cellular component. We found that hypoxic human mammary CAFs promoted angiogenesis in CAF-endothelial cell cocultures in vitro. Mass spectrometry-based proteomic analysis of the CAF secretome unraveled that hypoxic CAFs contributed to blood vessel abnormalities by altering their secretion of various pro- and anti-angiogenic factors...
February 5, 2019: Science Signaling
Shahar Ben-Shaul, Shira Landau, Uri Merdler, Shulamit Levenberg
Graft vascularization remains one of the most critical challenges facing tissue-engineering experts in their attempt to create thick transplantable tissues and organs. In vitro prevascularization of engineered tissues has been suggested to promote rapid anastomosis between the graft and host vasculatures; however, thrombotic events have been reported upon graft implantation. Here, we aimed to determine whether in vitro vessel maturation in transplantable grafts can accelerate vascular integration and graft perfusion and prevent thrombotic events in the grafts...
February 4, 2019: Proceedings of the National Academy of Sciences of the United States of America
Erhui Jiang, Zhi Xu, Meng Wang, Tinglin Yan, Chunming Huang, Xiaocheng Zhou, Qing Liu, Lin Wang, Yang Chen, Hui Wang, Ke Liu, Zhe Shao, Zhenjun Shang
Metabolic reprogramming is a hallmark of cancer. Stromal cells could function as providers of energy metabolites for tumor cells by undergoing the "reverse Warburg effect," but the mechanism has not been fully elucidated. The interaction between the tumoral microvesicles (TMVs) and stroma in the tumor microenvironment plays a critical role in facilitating cancer progression. In this study, we demonstrated a novel mechanism for the TMV-mediated glycometabolic reprogramming of stromal cells. After being incubated with TMVs, normal human gingival fibroblasts exhibited a phenotype switch to cancer-associated fibroblasts and underwent a degradation of caveolin 1 (CAV1) through the ERK1/2-activation pathway...
January 30, 2019: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Fátima Manzano-Núñez, María José Arámbul-Anthony, Amparo Galán Albiñana, Aranzazu Leal Tassias, Carlos Acosta Umanzor, Irene Borreda Gascó, Antonio Herrera, Jerónimo Forteza Vila, Deborah J Burks, Luke A Noon
Insulin provides important information to tissues about feeding behavior and energy status. Defective insulin signaling is associated with ageing, tissue dysfunction, and impaired wound healing. In the liver, insulin resistance leads to chronic damage and fibrosis, but it is unclear how tissue-repair mechanisms integrate insulin signals to coordinate an appropriate injury response or how they are affected by insulin resistance. In this study, we demonstrate that insulin resistance impairs local cellular crosstalk between the fibrotic stroma and bipotent adult liver progenitor cells (LPCs), whose paracrine interactions promote epithelial repair and tissue remodeling...
January 29, 2019: PLoS Biology
Monika Lodyga, Elizabeth Cambridge, Henna M Karvonen, Pardis Pakshir, Brian Wu, Stellar Boo, Melanie Kiebalo, Riitta Kaarteenaho, Michael Glogauer, Mohit Kapoor, Kjetil Ask, Boris Hinz
Macrophages contribute to the activation of fibroblastic cells into myofibroblasts, which secrete collagen and contract the collagen matrix to acutely repair injured tissue. Persistent myofibroblast activation leads to the accumulation of fibrotic scar tissue that impairs organ function. We investigated the key processes that turn acute beneficial repair into destructive progressive fibrosis. We showed that homotypic cadherin-11 interactions promoted the specific binding of macrophages to and persistent activation of profibrotic myofibroblasts...
January 15, 2019: Science Signaling
Miki Nishio, Yousuke Miyachi, Junji Otani, Shoji Tane, Hirofumi Omori, Fumihito Ueda, Hideru Togashi, Takehiko Sasaki, Tak Wah Mak, Kazuwa Nakao, Yasuyuki Fujita, Hiroshi Nishina, Tomohiko Maehama, Akira Suzuki
Cell competition is involved in mammalian embryogenesis and tumor elimination and progression. It was previously shown that, whereas NIH3T3 mouse fibroblasts expressing high levels of the yes-associated protein 1(YAP1) target TEA domain family (TEAD) transcription factors become "winners" in cell competitions, Madin-Darby canine kidney cells expressing activated YAP1 become "losers" and are eliminated from culture monolayers. Thus, YAP1's role in cell competitions is clearly context dependent...
January 14, 2019: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Tao Zhou, Wei Wang, Yasen Aimaiti, Xin Jin, Zhixin Chen, Liang Chen, Dewei Li
The widespread use of hepatocytes and cholangiocytes for regenerative medicine and tissue engineering is restricted by the limited number of hepatocytes and cholangiocytes; a simple and effective method for the expansion and differentiation of the hepatic progenitor cells (HPCs) is required. Recent studies demonstrated that mouse embryonic fibroblasts (MEFs) play an important role in supporting the proliferation of the mouse hepatic progenitor cells (mHPCs). However, the effect of direct and indirect coculture of MEFs with mHPCs on the differentiation of mHPCs is poorly studied...
January 2, 2019: Cytotechnology
Hae-Rim Kim, Bo-Mi Kim, Ji-Yeon Won, Kyung-Ann Lee, Hyun Myung Ko, Young Sun Kang, Sang-Heon Lee, Kyoung-Woon Kim
We investigated the immune-regulatory function of quercetin, in interleukin (IL)-17-produced osteoclastogenesis in rheumatoid arthritis (RA). RA fibroblasts-like synoviocytes (RA-FLS) were stimulated with IL-17, and the mRNA expression and secretion of receptor activator of nuclear factor kappa-B ligand (RANKL) were detected by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. CD14+ monocytes (osteoclast precursors) were stimulated with IL-17, RANKL, with/without quercetin, and tartrate-resistant acid phosphatase activity was evaluated to assess osteoclast differentiation...
February 2019: Journal of Medicinal Food
Stefano Castellani, Sante Di Gioia, Lorena di Toma, Massimo Conese
Chronic inflammation, oxidative stress, mucus plugging, airway remodeling, and respiratory infections are the hallmarks of the cystic fibrosis (CF) lung disease. The airway epithelium is central in the innate immune responses to pathogens colonizing the airways, since it is involved in mucociliary clearance, senses the presence of pathogens, elicits an inflammatory response, orchestrates adaptive immunity, and activates mesenchymal cells. In this review, we focus on cellular models of the human CF airway epithelium that have been used for studying mucus production, inflammatory response, and airway remodeling, with particular reference to two- and three-dimensional cultures that better recapitulate the native airway epithelium...
2018: Analytical Cellular Pathology (Amsterdam)
Tom S Chan, Hongbin Yu, Amanda Moore, Salman R Khetani, Donald Tweedie
Generating accurate in vitro intrinsic clearance data is an important aspect of predicting in vivo human clearance. Primary hepatocytes in suspension are routinely used to predict in vivo clearance; however, incubation times have typically been limited to 4-6 hours, which is not long enough to accurately evaluate the metabolic stability of slowly metabolized compounds. HepatoPac is a micropatterened hepatocyte-fibroblast coculture system that can be used for continuous incubations of up to 7 days. This study evaluated the ability of human HepatoPac to predict the in vivo clearance (CL) of 17 commercially available compounds with low to intermediate clearance (<12 ml/min/kg)...
January 2019: Drug Metabolism and Disposition: the Biological Fate of Chemicals
Sahar Biglari, Thi Y L Le, Richard P Tan, Steven G Wise, Alessandro Zambon, Gaia Codolo, Marina De Bernard, Majid Warkiani, Aaron Schindeler, Sina Naficy, Peter Valtchev, Ali Khademhosseini, Fariba Dehghani
Considerable progress has been made in the field of microfluidics to develop complex systems for modeling human skin and dermal wound healing processes. While microfluidic models have attempted to integrate multiple cell types and/or 3D culture systems, to date they have lacked some elements needed to fully represent dermal wound healing. This paper describes a cost-effective, multicellular microfluidic system that mimics the paracrine component of early inflammation close to normal wound healing. Collagen and Matrigel are tested as materials for coating and adhesion of dermal fibroblasts and human umbilical vein endothelial cells (HUVECs)...
December 4, 2018: Advanced Healthcare Materials
Heide-Katharina Bauer, Daniela Flesch, Stefan Walenta, Ronald E Unger, Roxana Schwab, Sandra Nezi-Cahn, Annette Hasenburg, Martin Heller, Walburgis Brenner
BACKGROUND: Our aim was to establish a reliable protocol for the isolation of primary human oral mucosa epithelial cells that ensured a successful, efficient and reproducible cell culture for the utilization in subsequent experiments and for tissue engineering purposes. Therefore, we analyzed and evaluated various isolation routines and cultivation protocols. Furthermore, we investigated the functionality of isolated epithelial cells for tissue engineering approaches in an in vitro mucosa equivalent cell culture model system...
December 1, 2018: Tissue Engineering. Part C, Methods
Shuang Chen, Andreas Giannakou, Sarah Wyman, Janet Gruzas, Jonathon Golas, Wenyan Zhong, Christine Loreth, Latha Sridharan, Ting-Ting Yamin, Marc Damelin, Kenneth G Geles
Tumorigenesis depends on intricate interactions between genetically altered tumor cells and their surrounding microenvironment. While oncogenic drivers in lung squamous carcinoma (LUSC) have been described, the role of stroma in modulating tissue architecture, particularly cell polarity, remains unclear. Here, we report the establishment of a 3D coculture system of LUSC epithelial cells with cancer-associated fibroblasts (CAFs) and extracellular matrix that together capture key components of the tumor microenvironment (TME)...
November 28, 2018: Proceedings of the National Academy of Sciences of the United States of America
Yuan Liu, Lu Gao, Xiaoyan Zhao, Sen Guo, Yuzhou Liu, Ran Li, Cui Liang, Ling Li, Jianzeng Dong, Lina Li, Haibo Yang
Saikosaponin A (SSA) is a triterpenoid saponin with many pharmacological activities, including anti-inflammatory and antioxidant effects. The effect of SSA on cardiac remodeling and fibrosis, however, remains unclear. Aortic banding surgery was used to establish a mouse cardiac remodeling and fibrosis model. Mice were subjected to an intraperitoneal (i.p.) injection of SSA (5 mg/kg/d or 40 mg/kg/d) 2 weeks after surgery for 28 days. As a result, SSA had limited effect on cardiac hypertrophy but decreased cardiac fibrosis remarkably...
2018: International Journal of Biological Sciences
Ton Schoenmaker, Fenne Wouters, Dimitra Micha, Tim Forouzanfar, Coen Netelenbos, E Marelise W Eekhoff, Nathalie Bravenboer, Teun J de Vries
Fibrodysplasia ossificans progressiva (FOP) is a genetic disease characterized by heterotopic ossification (HO). The disease is caused by a mutation in the activin receptor type 1 (ACVR1) gene that enhances this receptor's responsiveness to Activin-A. Binding of Activin-A to the mutated ACVR1 receptor induces osteogenic differentiation. Whether Activin-A also affects osteoclast formation in FOP is not known. Therefore we investigated its effect on the osteoclastogenesis-inducing potential of periodontal ligament fibroblasts (PLF) from teeth of healthy controls and patients with FOP...
November 11, 2018: Journal of Cellular Physiology
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