scholarly journals Usefulness of Mesenchymal Cell Lines for Bone and Cartilage Regeneration Research

2019 ◽  
Vol 20 (24) ◽  
pp. 6286 ◽  
Author(s):  
M. Piñeiro-Ramil ◽  
C. Sanjurjo-Rodríguez ◽  
R. Castro-Viñuelas ◽  
S. Rodríguez-Fernández ◽  
I.M. Fuentes-Boquete ◽  
...  
Keyword(s):  
Disease Modeling ◽  
Cartilage Regeneration ◽  
Mesenchymal Cell ◽  
Original Research ◽  
Full Potential ◽  
Primary Cells ◽  
Parental Phenotype ◽  
In Vitro Repair ◽  
And Cartilage

The unavailability of sufficient numbers of human primary cells is a major roadblock for in vitro repair of bone and/or cartilage, and for performing disease modelling experiments. Immortalized mesenchymal stromal cells (iMSCs) may be employed as a research tool for avoiding these problems. The purpose of this review was to revise the available literature on the characteristics of the iMSC lines, paying special attention to the maintenance of the phenotype of the primary cells from which they were derived, and whether they are effectively useful for in vitro disease modeling and cell therapy purposes. This review was performed by searching on Web of Science, Scopus, and PubMed databases from 1 January 2015 to 30 September 2019. The keywords used were ALL = (mesenchymal AND (“cell line” OR immortal*) AND (cartilage OR chondrogenesis OR bone OR osteogenesis) AND human). Only original research studies in which a human iMSC line was employed for osteogenesis or chondrogenesis experiments were included. After describing the success of the immortalization protocol, we focused on the iMSCs maintenance of the parental phenotype and multipotency. According to the literature revised, it seems that the maintenance of these characteristics is not guaranteed by immortalization, and that careful selection and validation of clones with particular characteristics is necessary for taking advantage of the full potential of iMSC to be employed in bone and cartilage-related research.

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.

scholarly journals Progress and Applications of Polyphosphate in Bone and Cartilage Regeneration

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Yan Wang ◽  
Min Li ◽  
Pei Li ◽  
Haijun Teng ◽  
Dehong Fan ◽  
...  
Keyword(s):  
Cartilage Regeneration ◽  
High Energy ◽  
Cartilage Defects ◽  
Artificial Bone ◽  
Cellular Mechanisms ◽  
Inorganic Polyphosphate ◽  
Morphogenetic Effects ◽  
And Cartilage

Patients with bone and cartilage defects due to infection, tumors, and trauma are quite common. Repairing bone and cartilage defects is thus a major problem for clinicians. Autologous and artificial bone transplantations are associated with many challenges, such as limited materials and immune rejection. Bone and cartilage regeneration has become a popular research topic. Inorganic polyphosphate (polyP) is a widely occurring biopolymer with high-energy phosphoanhydride bonds that exists in organisms from bacteria to mammals. Much data indicate that polyP acts as a regulator of gene expression in bone and cartilage tissues and exerts morphogenetic effects on cells involved in bone and cartilage formation. Exposure of these cells to polyP leads to the increase of cytokines that promote the differentiation of mesenchymal stem cells into osteoblasts, accelerates the osteoblast mineralization process, and inhibits the differentiation of osteoclast precursors to functionally active osteoclasts. PolyP-based materials have been widely reported in in vivo and in vitro studies. This paper reviews the current cellular mechanisms and material applications of polyP in bone and cartilage regeneration.

Arg-Gly-Asp Peptide-Functionalized Superparamagnetic γ-Fe2O3 Nanoparticles Enhance Osteoblast Migration In Vitro

2020 ◽  
Vol 20 (10) ◽  
pp. 6173-6179
Author(s):  
Xue Liu ◽  
Xiao-Ling Yang ◽  
Qiao Hu ◽  
Mao-Shi Liu ◽  
Tao Peng ◽  
...  
Keyword(s):  
Magnetic Field ◽  
Cell Therapy ◽  
Bone Repair ◽  
Cartilage Regeneration ◽  
Uniform Size ◽  
Low Cytotoxicity ◽  
The Magnetic Field ◽  
And Cartilage ◽  
Osteoblast Migration

Making osteoblast migration manageably target to injury sites has been the key challenging in cell therapy for bone and cartilage regeneration. Superparamagnetic materials, the magnetic guide for cell migration, have been applied to increase cell retention. However, additional targeting modifications are still needed to accelerate the low uptake efficiency and moving speed. Arg-Gly-Asp peptide (RGD)-functionalized magnetic nanoparticles showed cutting-edge competence in cell differentiation control and targeted drug delivery. However, more evidence was required to corroborate its role in osteoblast migration in bone repair. In the present study, RGD-modified γ-Fe2O3 nanoparticles (RGD-Fe2O3 NPs) were prefabricated with the grafting ratio of 33.3–37.4%. The RGD-Fe2O3 NPs unveiled excellent water dispersibility with uniform size distribution at 5–6 nm and negligibly low cytotoxicity. As a result, MC3T3-E1 osteoblasts treated with RGD-Fe2O3 NPs boosted its migration speed in a magnetic field compared with those incubated with unmodified Fe2O3 NPs. Furthermore, osteoblasts treated with RGD-Fe2O3 NPs exhibited more Fe uptake. The results exposed the fact that RGD-mediated specific cellular uptake presented higher efficiency than the non-RGD-mediated one, resulting from a stronger superparamagnetic force between the labeled cells and the magnetic field. These findings indicate that the RGD-functionalized Fe2O3 NPs can promote osteoblast migration in the magnetic field, providing a promising strategy in magnet-guided cell therapy for bone and cartilage regeneration.

scholarly journals Ethanol treatment of nanoPGA/PCL composite scaffolds enhances human chondrocyte development in the cellular microenvironment of tissue-engineered auricle constructs

PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0253149
Author(s):  
Narihiko Hirano ◽  
Hirohisa Kusuhara ◽  
Yu Sueyoshi ◽  
Takeshi Teramura ◽  
Ananth Murthy ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cartilage Regeneration ◽  
Cellular Adhesion ◽  
Ethanol Treatment ◽  
Cellular Microenvironment ◽  
Composite Scaffolds ◽  
Expression Levels ◽  
Cell Scaffold ◽  
And Cartilage

A major obstacle for tissue engineering ear-shaped cartilage is poorly developed tissue comprising cell-scaffold constructs. To address this issue, bioresorbable scaffolds of poly-ε-caprolactone (PCL) and polyglycolic acid nanofibers (nanoPGA) were evaluated using an ethanol treatment step before auricular chondrocyte scaffold seeding, an approach considered to enhance scaffold hydrophilicity and cartilage regeneration. Auricular chondrocytes were isolated from canine ears and human surgical samples discarded during otoplasty, including microtia reconstruction. Canine chondrocytes were seeded onto PCL and nanoPGA sheets either with or without ethanol treatment to examine cellular adhesion in vitro. Human chondrocytes were seeded onto three-dimensional bioresorbable composite scaffolds (PCL with surface coverage of nanoPGA) either with or without ethanol treatment and then implanted into athymic mice for 10 and 20 weeks. On construct retrieval, scanning electron microscopy showed canine auricular chondrocytes seeded onto ethanol-treated scaffolds in vitro developed extended cell processes contacting scaffold surfaces, a result suggesting cell-scaffold adhesion and a favorable microenvironment compared to the same cells with limited processes over untreated scaffolds. Adhesion of canine chondrocytes was statistically significantly greater (p ≤ 0.05) for ethanol-treated compared to untreated scaffold sheets. After implantation for 10 weeks, constructs of human auricular chondrocytes seeded onto ethanol-treated scaffolds were covered with glossy cartilage while constructs consisting of the same cells seeded onto untreated scaffolds revealed sparse connective tissue and cartilage regeneration. Following 10 weeks of implantation, RT-qPCR analyses of chondrocytes grown on ethanol-treated scaffolds showed greater expression levels for several cartilage-related genes compared to cells developed on untreated scaffolds with statistically significantly increased SRY-box transcription factor 5 (SOX5) and decreased interleukin-1α (inflammation-related) expression levels (p ≤ 0.05). Ethanol treatment of scaffolds led to increased cartilage production for 20- compared to 10-week constructs. While hydrophilicity of scaffolds was not assessed directly in the present findings, a possible factor supporting the summary data is that hydrophilicity may be enhanced for ethanol-treated nanoPGA/PCL scaffolds, an effect leading to improvement of chondrocyte adhesion, the cellular microenvironment and cartilage regeneration in tissue-engineered auricle constructs.

scholarly journals In Vitro Cartilage Regeneration with a Three-Dimensional Polyglycolic Acid (PGA) Implant in a Bovine Cartilage Punch Model

2021 ◽  
Vol 22 (21) ◽  
pp. 11769
Author(s):  
Victoria Horbert ◽  
Long Xin ◽  
Peter Föhr ◽  
René Huber ◽  
Rainer H. Burgkart ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cartilage Regeneration ◽  
Polyglycolic Acid ◽  
Cartilage Defects ◽  
Human Articular Cartilage ◽  
Chain Reactions ◽  
Bovine Cartilage Punch Model ◽  
Push Out ◽  
And Cartilage

Resorbable polyglycolic acid (PGA) chondrocyte grafts are clinically established for human articular cartilage defects. Long-term implant performance was addressed in a standardized in vitro model. PGA implants (+/− bovine chondrocytes) were placed inside cartilage rings punched out of bovine femoral trochleas (outer Ø 6 mm; inner defect Ø 2 mm) and cultured for 84 days (12 weeks). Cartilage/PGA hybrids were subsequently analyzed by histology (hematoxylin/eosin; safranin O), immunohistochemistry (aggrecan, collagens 1 and 2), protein assays, quantitative real-time polymerase chain reactions, and implant push-out force measurements. Cartilage/PGA hybrids remained vital with intact matrix until 12 weeks, limited loss of proteoglycans from “host” cartilage or cartilage–PGA interface, and progressively diminishing release of proteoglycans into the supernatant. By contrast, the collagen 2 content in cartilage and cartilage–PGA interface remained approximately constant during culture (with only little collagen 1). Both implants (+/− cells) displayed implant colonization and progressively increased aggrecan and collagen 2 mRNA, but significantly decreased push-out forces over time. Cell-loaded PGA showed significantly accelerated cell colonization and significantly extended deposition of aggrecan. Augmented chondrogenic differentiation in PGA and cartilage/PGA-interface for up to 84 days suggests initial cartilage regeneration. Due to the PGA resorbability, however, the model exhibits limitations in assessing the “lateral implant bonding”.

scholarly journals PuraMatrix allows differentiation of a broad repertoire of neural and mesenchymal phenotypes from trunk neural crest

2020 ◽  
Vol 64 (7-8-9) ◽  
pp. 433-443
Author(s):  
Clarissa R. Taufer ◽  
Monica A. Rodrigues-Da-Silva ◽  
Giordano W. Calloni
Keyword(s):  
Neural Crest ◽  
Mesenchymal Cell ◽  
Cell Types ◽  
Full Potential ◽  
Self Renewal ◽  
Mesenchymal Differentiation ◽  
Trunk Neural Crest ◽  
And Control

The neural crest (NC) is a transitory embryonic structure of vertebrates that gives rise to an astonishing variety of derivatives, encompassing both neural and mesenchymal cell types. Neural crest cells (NCCs) are an excellent model to study how environmental factors modulate features such as cell multipotentiality and differentiation. Tests with multifunctional substrates that allow NCCs to express their full potential, while promoting cell subcloning, are needed to advance knowledge about NCC self-renewal and to foster future biotechnological approaches. Here we show that a self-assembled peptide named PuraMatrixTM is an excellent substrate that allows the differentiation of NCCs based on the identification of seven different cell types. Depending on the PuraMatrixTM concentration employed, different frequencies and quantities of a given cell type were obtained. It is noteworthy that an enormous quantity and diversity of mesenchymal phenotypes, such as chondrocytes, could be observed. The quantity of adipocytes and osteocytes also increased with the use of mesenchymal differentiation factors (MDF), but PuraMatrixTM alone can support the appearance of these mesenchymal cell types. PuraMatrixTM will promote advances in studies related to multipotentiality, self-renewal and control of NCC differentiation, since it is an extremely simple and versatile material which can be employed for both in vivo and in vitro experiments.

Culture Expanded Canine Mesenchymal Stem Cells Possess Osteochondrogenic Potential in Vivo and in Vitro

1997 ◽  
Vol 6 (2) ◽  
pp. 125-134 ◽  
Author(s):  
S. Kadiyala ◽  
R. G. Young ◽  
M. A. Thiede ◽  
S. P. Bruder
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Morphological Characteristics ◽  
Cartilage Regeneration ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Human Mscs ◽  
And Cartilage

Mesenchymal Stem Cells (MSCs) possessing the capacity to differentiate into various cell types such as osteoblasts, chondrocytes, myoblasts, and adipocytes have been previously isolated from the marrow and periosteum of human, murine, lapine, and avian species. This study documents the existence of similar multipotential stem cells in canine marrow. The cells were isolated from marrow aspirates using a modification of techniques previously established for human MSCs (hMSCs), and found to possess similar growth and morphological characteristics, as well as osteochondrogenic potential in vivo and in vitro. On the basis of these results, the multipotential cells that were isolated and culture expanded are considered to be canine MSCs (cMSCs). The occurrence of cMSCs in the marrow was determined to be one per 2.5 × 104 nucleated cells. After enrichment of the cMSCs by centrifugation on a Per-coll cushion, the cells were cultivated in selected lots of serum. Like the hMSCs, cMSCs grew as colonies in primary culture and on replating, grew as a monolayer culture with very uniform spindle morphology. The population doubling time for these cMSCs was approximately 2 days. The morphology and the growth kinetics of the cMSCs were retained following repeated passaging. The osteogenic phenotype could be induced in the cMSC cultures by the addition of a synthetic glucocorticoid, dexamethasone. In these osteogenic cultures, alkaline phosphatase activity was elevated up to 10-fold, and mineralized matrix production was evident. When cMSCs were loaded onto porous ceramics and implanted in autologous canine or athymic murine hosts, copious amounts of bone and cartilage were formed in the pores of the implants. The MSC-mediated osteogenesis obtained by the implantation of the various MSC-loaded matrix combinations is the first evidence of osteogenesis in a canine model by implantation of culture expanded autologous stem cells. The identification and isolation of cMSCs now makes it feasible to pursue preclinical models of bone and cartilage regeneration in canine hosts.

scholarly journals Mechano-modulatory synthetic niches for liver organoid derivation

2019 ◽  
Author(s):  
Giovanni Sorrentino ◽  
Saba Rezakhani ◽  
Ece Yildiz ◽  
Sandro Nuciforo ◽  
Markus H. Heim ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Disease Modeling ◽  
Three Dimensional ◽  
Real Life ◽  
Matrix Stiffness ◽  
Primary Cells ◽  
Proliferative Capacity ◽  
Liver Organoids ◽  
Recent Demonstration

AbstractThe recent demonstration that primary cells from the liver can be expanded in vitro as organoids holds enormous promise for regenerative medicine and disease modeling1–5. The use of three-dimensional (3D) cultures based on ill-defined and potentially immunogenic matrices, however, hampers the translation of liver organoid technology into real-life applications6. We here used chemically defined hydrogels for the efficient derivation of both mouse and human hepatic organoids. Organoid growth was found to be highly stiffness-sensitive and dependent on yes-associated protein 1 (YAP) activity. However, in contrast to intestinal organoids7, YAP-mediated stiffness sensitivity was independent of acto-myosin contractility, requiring instead activation of the Src family of kinases (SFKs). Aberrant matrix stiffness on the other hand led to a shift in the progenitor phenotype, resulting in compromised proliferative capacity. Finally, we demonstrate the unprecedented establishment of biopsy-derived human liver organoids without the use of animal components at any step of the process. Our approach thus opens up exciting perspectives for the establishment of protocols for liver organoid-based regenerative medicine.

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