scholarly journals Microvalve bioprinting of MSC-Chondrocyte Co-Cultures

2021 ◽  
Author(s):  
Joseph Dudman ◽  
Ana Marina Ferreira ◽  
Piergiorgio Gentile ◽  
Xiao Wang ◽  
Kenneth Dalgarno
Keyword(s):  
High Throughput Screening ◽  
Cellular Therapy ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Proof Of Concept ◽  
Concept Study ◽  
Indirect Immunofluorescence Staining ◽  
Chondrocyte Implantation ◽  
Tissue Models

Recent improvements within the fields of high-throughput screening and 3D tissue culture have provided the possibility of developing in vitro micro-tissue models that can be used to study diseases and screen potential new therapies. This paper reports a proof of concept study on the use of microvalve-based bioprinting to create laminar MSC-chondrocyte co-cultures as an in vitro model of autologous chondrocyte implantation (ACI), an established cellular therapy for osteoarthritis. Microvalve-based bioprinting uses microvalves to deposit cells suspended in a liquid in a consistent and repeatable manner. In this case MSCs and chondrocytes have been sequentially deposited into an insert based transwell system in order to create a laminar co-culture, with variations in the ratios of the cell types used to investigate the potential for MSCs to stimulate improved repair. Histological and indirect immunofluorescence staining revealed the formation of dense tissue structures within the chondrocyte and MSC-chondrocyte cell co-cultures, alongside the establishment of a proliferative region at the base of the tissue. No stimulatory or inhibitory effect in terms of ECM production was observed through the introduction of MSCs, although the potential for an immunomodulatory benefit remains. This proof-of-concept study therefore provides a novel method to enable the scalable production of therapeutically relevant micro-tissue models that can be used for in vitro research to optimise ACI procedures.

scholarly journals Microvalve Bioprinting of MSC-Chondrocyte Co-Cultures

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3329
Author(s):  
Joseph Dudman ◽  
Ana Marina Ferreira ◽  
Piergiorgio Gentile ◽  
Xiao Wang ◽  
Kenneth Dalgarno
Keyword(s):  
High Throughput Screening ◽  
Cell Types ◽  
Inhibitory Effect ◽  
Small Scale ◽  
Indirect Immunofluorescence Staining ◽  
Dense Tissue ◽  
Novel Method ◽  
Tissue Models ◽  
Transwell System

Recent improvements within the fields of high-throughput screening and 3D tissue culture have provided the possibility of developing in vitro micro-tissue models that can be used to study diseases and screen potential new therapies. This paper reports a proof-of-concept study on the use of microvalve-based bioprinting to create laminar MSC-chondrocyte co-cultures to investigate whether the use of MSCs in ACI procedures would stimulate enhanced ECM production by chondrocytes. Microvalve-based bioprinting uses small-scale solenoid valves (microvalves) to deposit cells suspended in media in a consistent and repeatable manner. In this case, MSCs and chondrocytes have been sequentially printed into an insert-based transwell system in order to create a laminar co-culture, with variations in the ratios of the cell types used to investigate the potential for MSCs to stimulate ECM production. Histological and indirect immunofluorescence staining revealed the formation of dense tissue structures within the chondrocyte and MSC-chondrocyte cell co-cultures, alongside the establishment of a proliferative region at the base of the tissue. No stimulatory or inhibitory effect in terms of ECM production was observed through the introduction of MSCs, although the potential for an immunomodulatory benefit remains. This study, therefore, provides a novel method to enable the scalable production of therapeutically relevant micro-tissue models that can be used for in vitro research to optimise ACI procedures.

scholarly journals Tenovin-6 impairs autophagy by inhibiting autophagic flux

2017 ◽  
Vol 8 (2) ◽  
pp. e2608-e2608 ◽  
Author(s):  
Hongfeng Yuan ◽  
Brandon Tan ◽  
Shou-Jiang Gao
Keyword(s):  
Cell Types ◽  
Inhibitory Effect ◽  
Lymphocytic Leukemia ◽  
Autophagic Flux ◽  
Dependent Manner ◽  
Autophagy Pathway ◽  
Sarcoma Cells ◽  
Regulatory Functions

Abstract Tenovin-6 has attracted significant interest because it activates p53 and inhibits sirtuins. It has anti-neoplastic effects on multiple hematopoietic malignancies and solid tumors in both in vitro and in vivo studies. Tenovin-6 was recently shown to impair the autophagy pathway in chronic lymphocytic leukemia cells and pediatric soft tissue sarcoma cells. However, whether tenovin-6 has a general inhibitory effect on autophagy and whether there is any involvement with SIRT1 and p53, both of which are regulators of the autophagy pathway, remain unclear. In this study, we have demonstrated that tenovin-6 increases microtubule-associated protein 1 light chain 3 (LC3-II) level in diverse cell types in a time- and dose-dependent manner. Mechanistically, the increase of LC3-II by tenovin-6 is caused by inhibition of the classical autophagy pathway via impairing lysosomal function without affecting the fusion between autophagosomes and lysosomes. Furthermore, we have revealed that tenovin-6 activation of p53 is cell type dependent, and tenovin-6 inhibition of autophagy is not dependent on its regulatory functions on p53 and SIRT1. Our results have shown that tenovin-6 is a potent autophagy inhibitor, and raised the precaution in interpreting results where tenovin-6 is used as an inhibitor of SIRT1.

scholarly journals Electrophysiological Analysis of Brain Organoids: Current Approaches and Advancements

2021 ◽  
Vol 14 ◽  
Author(s):  
Austin P. Passaro ◽  
Steven L. Stice
Keyword(s):  
High Throughput Screening ◽  
Disease Modeling ◽  
Cell Types ◽  
Full Potential ◽  
Two Dimensional ◽  
Physiological Relevance ◽  
Human Cell Cultures ◽  
Brain Organoid ◽  
Electrophysiological Methods

Brain organoids, or cerebral organoids, have become widely used to study the human brain in vitro. As pluripotent stem cell-derived structures capable of self-organization and recapitulation of physiological cell types and architecture, brain organoids bridge the gap between relatively simple two-dimensional human cell cultures and non-human animal models. This allows for high complexity and physiological relevance in a controlled in vitro setting, opening the door for a variety of applications including development and disease modeling and high-throughput screening. While technologies such as single cell sequencing have led to significant advances in brain organoid characterization and understanding, improved functional analysis (especially electrophysiology) is needed to realize the full potential of brain organoids. In this review, we highlight key technologies for brain organoid development and characterization, then discuss current electrophysiological methods for brain organoid analysis. While electrophysiological approaches have improved rapidly for two-dimensional cultures, only in the past several years have advances been made to overcome limitations posed by the three-dimensionality of brain organoids. Here, we review major advances in electrophysiological technologies and analytical methods with a focus on advances with applicability for brain organoid analysis.

A proof-of-concept study: evaluating the applicability of high-throughput screening data and read-across tools for food relevant chemicals

2018 ◽  
Vol 295 ◽  
pp. S142
Author(s):  
J.L. Nguyen ◽  
A. Maier ◽  
J. Ovesen ◽  
N. Kleinstreuer ◽  
R. Judson ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Proof Of Concept ◽  
Concept Study

scholarly journals Autologous Bone Marrow Mononuclear Cell Therapy for Autism: An Open Label Proof of Concept Study

2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Alok Sharma ◽  
Nandini Gokulchandran ◽  
Hemangi Sane ◽  
Anjana Nagrajan ◽  
Amruta Paranjape ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cellular Therapy ◽  
Autologous Bone Marrow ◽  
Bone Marrow Mononuclear Cell ◽  
Clinical Effects ◽  
Proof Of Concept ◽  
Open Label ◽  
Global Improvement ◽  
Concept Study ◽  
Autologous Bone

Cellular therapy is an emerging therapeutic modality with a great potential for the treatment of autism. Recent findings show that the major underlying pathogenetic mechanisms of autism are hypoperfusion and immune alterations in the brain. So conceptually, cellular therapy which facilitates counteractive processes of improving perfusion by angiogenesis and balancing inflammation by immune regulation would exhibit beneficial clinical effects in patients with autism. This is an open label proof of concept study of autologous bone marrow mononuclear cells (BMMNCs) intrathecal transplantation in 32 patients with autism followed by multidisciplinary therapies. All patients were followed up for 26 months (mean 12.7). Outcome measures used were ISAA, CGI, and FIM/Wee-FIM scales. Positron Emission Tomography-Computed Tomography (PET-CT) scan recorded objective changes. Out of 32 patients, a total of 29 (91%) patients improved on total ISAA scores and 20 patients (62%) showed decreased severity on CGI-I. The difference between pre- and postscores was statistically significant (P<0.001) on Wilcoxon matched-pairs signed rank test. On CGI-II 96% of patients showed global improvement. The efficacy was measured on CGI-III efficacy index. Few adverse events including seizures in three patients were controlled with medications. The encouraging results of this leading clinical study provide future directions for application of cellular therapy in autism.

In vitro effects of mechanical stimulation and photobiomodulation on osteoblastic cell function: A proof of concept study

2017 ◽  
Vol 27 (1) ◽  
pp. 29-41 ◽  
Author(s):  
Rochaya Chintavalakorn ◽  
Anak Khantachawana ◽  
Kwanchanok Viravaidya-Pasuwat ◽  
Peerapong Santiwong ◽  
Rudee Surarit
Keyword(s):  
Mechanical Stimulation ◽  
Cell Function ◽  
Osteoblastic Cell ◽  
Proof Of Concept ◽  
Concept Study ◽  
In Vitro Effects

scholarly journals CARPOOL: A library-based platform to rapidly identify next generation chimeric antigen receptors

2021 ◽  
Author(s):  
Taeyoon Kyung ◽  
Khloe S Gordon ◽  
Caleb R Perez ◽  
Patrick V Holec ◽  
Azucena Ramos ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Cell Types ◽  
Tumor Control ◽  
Antigen Receptors ◽  
Cell Therapies ◽  
Promising Tool ◽  
Signaling Domain ◽  
Screening Platform

CD19-targeted CAR therapies have successfully treated B cell leukemias and lymphomas, but many responders later relapse or experience toxicities. CAR intracellular domains (ICDs) are key to converting antigen recognition into anti-tumor effector functions. Despite the many possible immune signaling domain combinations that could be included in CARs, almost all CARs currently rely upon CD3𝛇, CD28, and/or 4-1BB signaling. To explore the signaling potential of CAR ICDs, we generated a library of 700,000 CD19 CAR molecules with diverse signaling domains and developed a high throughput screening platform to enable optimization of CAR signaling for anti-tumor functions. Our strategy identifies CARs with novel signaling domain combinations that elicit distinct T cell behaviors from a clinically available CAR, including enhanced proliferation and persistence, lower exhaustion, potent cytotoxicity in an in vitro tumor rechallenge condition, and comparable tumor control in vivo. This approach is readily adaptable to numerous disease models, cell types, and selection conditions, making it a promising tool for rapidly improving adoptive cell therapies and expanding their utility to new disease indications.

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