scholarly journals Assessment of TGF-β3 on production of aggrecan by human articular chondrocytes in pellet culture system

2014 ◽  
Vol 3 (1) ◽  
pp. 54 ◽  
Author(s):  
Batool Hashemibeni ◽  
Azadeh Kabiri ◽  
Hossein Rabbani ◽  
Saeed Zamani ◽  
Ebrahim Esfandiari ◽  
...  
Keyword(s):  
Culture System ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
Pellet Culture

scholarly journals Adult human articular chondrocytes in a microcarrier-based culture system: expansion and redifferentiation

2010 ◽  
Vol 29 (4) ◽  
pp. 539-546 ◽  
Author(s):  
Karsten Schrobback ◽  
Travis Jacob Klein ◽  
Michael Schuetz ◽  
Zee Upton ◽  
David Ian Leavesley ◽  
...  
Keyword(s):  
Culture System ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
System Expansion ◽  
Adult Human

Rapid and Large-Scale Formation of Chondrocyte Aggregates by Rotational Culture

2003 ◽  
Vol 12 (5) ◽  
pp. 475-479 ◽  
Author(s):  
Katsuko S. Furukawa ◽  
Hideyuki Suenaga ◽  
Kenshi Toita ◽  
Akiko Numata ◽  
Junzo Tanaka ◽  
...  
Keyword(s):  
Culture System ◽  
Large Scale ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Cell Interaction ◽  
Collagen Type I ◽  
Cell Contact ◽  
Type I ◽  
Pellet Culture ◽  
Cell Cell

Chondrocytes in articular cartilage synthesize collagen type II and large sulfated proteoglycans, whereas the same cells cultured in monolayer (2D) dedifferentiate into fibroblastic cells and express collagen type I and small proteoglycans. On the other hand, a pellet culture system was developed as a method for preventing the phenotypic modulation of chondrocytes and promoting the redifferentiation of dedifferentiated ones. Because the pellet culture system forms only one cell aggregate each tube by a centrifugator, the pellet could not be applied to produce a tissue-engineered cartilage. Therefore, we tried to form chondrocyte aggregates by a rotational culture, expecting to form a large number of aggregates at once. In order to increase cell–cell interactions and decrease chondrocyte–material interaction, dishes with low retention of protein adsorption and cell adhesiveness were used. In addition, rotational shaking of the dish including cells was attempted to increase the cell–cell interaction. The shaking speed was set at 80 rpm, so the cells would be distributed in the center of the dish to augment the frequency of cell–cell contact. Under these conditions, bovine articular chondrocytes started aggregating in a few hours. At 24–36 h of rotational culture, aggregates with smooth surfaces were observed. Parameters such as increase of culture time and addition of TGF-β controlled diameters of the aggregates. There were many fusiform cells at the periphery of the aggregates, where the cells tended to form a multilayered zone in cross sections. In addition, lacune-like structure, which was almost the same as pellet culture, was observed. It was found that the internal structure of the aggregates was similar to that of pellets reported previously. Therefore, the aggregates formed by a rotational culture could become an essential component to make tissue-engineered artificial cartilage.

scholarly journals Human Articular Chondrocytes Retain Their Phenotype in Sustained Hypoxia While Normoxia Promotes Their Immunomodulatory Potential

Cartilage ◽  
2018 ◽  
Vol 10 (4) ◽  
pp. 467-479 ◽  
Author(s):  
Claire Mennan ◽  
John Garcia ◽  
Helen McCarthy ◽  
Sharon Owen ◽  
Jade Perry ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Quantitative Polymerase Chain Reaction ◽  
Growth Factor Receptor ◽  
Interferon Γ ◽  
Human Articular Chondrocytes ◽  
Pellet Culture ◽  
3 Dimensional ◽  
Hypoxic Conditions ◽  
Chondrogenic Phenotype ◽  
Immunomodulatory Potential

Objective To assess the phenotype of human articular chondrocytes cultured in normoxia (21% O2) or continuous hypoxia (2% O2). Design Chondrocytes were extracted from patients undergoing total knee replacement ( n = 5) and cultured in ~21% (normoxic chondrocytes, NC) and 2% (hypoxic chondrocytes, HC) oxygen in both monolayer and 3-dimensional (3D) pellet culture and compared with freshly isolated chondrocytes (FC). Cells were assessed by flow cytometry for markers indicative of mesenchymal stromal cells (MSCs), chondrogenic-potency and dedifferentiation. Chondrogenic potency and immunomodulatory gene expression was assessed in NC and HC by reverse transcription quantitative polymerase chain reaction. Immunohistochemistry was used to assess collagen II production following 3D pellet culture. Results NC were positive (>97%, n = 5) for MSC markers, CD73, CD90, and CD105, while HC demonstrated <90% positivity ( n = 4) and FC ( n = 5) less again (CD73 and CD90 <20%; CD105 <40%). The markers CD166 and CD151, indicative of chondrogenic de-differentiation, were significantly higher on NC compared with HC and lowest on FC. NC also produced the highest levels of CD106 and showed the greatest levels of IDO expression, following interferon-γ stimulation, indicating immunomodulatory potential. NC produced the highest levels of CD49c (>60%) compared with HC and FC in which production was <2%. Hypoxic conditions upregulated expression of SOX9, frizzled-related protein ( FRZB), fibroblast growth factor receptor 3 ( FGFR3), and collagen type II ( COL2A1) and downregulated activin receptor-like kinase 1 ( ALK1) in 3 out of 4 patients compared with normoxic conditions for monolayer cells. Conclusions Hypoxic conditions encourage retention of a chondrogenic phenotype with some immunomodulatory potential, whereas normoxia promotes dedifferentiation of chondrocytes toward an MSC phenotype with loss of chondrogenic potency but enhanced immunomodulatory capacity.

scholarly journals Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

2011 ◽  
Vol 347 (3) ◽  
pp. 649-663 ◽  
Author(s):  
Karsten Schrobback ◽  
Travis Jacob Klein ◽  
Ross Crawford ◽  
Zee Upton ◽  
Jos Malda ◽  
...  
Keyword(s):  
Culture System ◽  
In Vitro Propagation ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes

Human Umbilical Cord Mesenchymal Stem Cells Induce into Chondrocytes in 3D Cultured System

2013 ◽  
Vol 749 ◽  
pp. 198-205
Author(s):  
Li Yu ◽  
Jing Liu ◽  
Chao Xu ◽  
Er Mei Luo ◽  
Ming Qiao Tang
Keyword(s):  
Real Time ◽  
Alcian Blue ◽  
Culture System ◽  
Type Ii Collagen ◽  
Human Umbilical Cord ◽  
Type Ii ◽  
Pellet Culture ◽  
Hla Dr

Objective: To investigate a better method of inducing hUC-MSCs into chondrocytes in different culture system in vitro. Method: hUC-MSCs were isolated and cultured by tissue block culture, and the cells surface antigens were identified by flow cytometry, hUC-MSCs were cultured with chondrogenic media and stained with Alcian Blue. The production of matrix was estimated from the determination of hydroxyproline content and Alcian Blue method. Expressions of glycosaminoglycan (GAG), type II collagen and Sox-9 were assayed by real-time fluorescence quantitative PCR. Results: The cultured hUC-MSCs phenotype was CD105+/CD29+/CD44+/ CD31-/CD34-/ CD40-/CD45-/HLA-DR-. hUC-MSCs weakly expressed chondrocyte marker, which strongly expressed GAG and type II collagen after chondrogenic induction, and the cells were incubated in pellet culture with higher expression. Real-time PCR results demonstrated that chondrogenic induction cells were expressed GAG, type II collagen and Sox-9, and the cells were incubated in pellet culture with higher expression. Conclusion: hUC-MSCs incubated in pellet culture is more conducive to differentiate into chondrocytes than those cultured in monolayer culture system.

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