A low temperature method of isolating normal human articular chondrocytes

Hidvégi, Norbert; Sales, Kevin M.; Izadi, Dena; Ong, Juling; Kellam, Paul; Eastwood, DM; Butler, Peter E. M.

doi:10.1016/j.joca.2005.08.007

Cited by 25 publications

(20 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previous studies have investigated temperature modulation [31], human serum supplementation [32], and the use of ascorbic acid and NaCl in perfusion bioreactor systems to enhance cell isolation protocols [14]. While these approaches are highly innovative and could add significantly to advances in GMP biomanufacturing, for large scale isolation and tissue engineering approaches, short and simple protocols are desirable for clinical translation.…”

Section: Discussionmentioning

confidence: 99%

Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

Vedicherla

Buckley

2017

BioMed Research International

View full text Add to dashboard Cite

Laboratory based processing and expansion to yield adequate cell numbers had been the standard in Autologous Disc Chondrocyte Transplantation (ADCT), Allogeneic Juvenile Chondrocyte Implantation (NuQu®), and Matrix-Induced Autologous Chondrocyte Implantation (MACI). Optimizing cell isolation is a key challenge in terms of obtaining adequate cell numbers while maintaining a vibrant cell population capable of subsequent proliferation and matrix elaboration. However, typical cell yields from a cartilage digest are highly variable between donors and based on user competency. The overall objective of this study was to optimize chondrocyte isolation from cartilaginous nasal tissue through modulation of enzyme concentration exposure (750 and 3000 U/ml) and incubation time (1 and 12 h), combined with physical agitation cycles, and to assess subsequent cell viability and matrix forming capacity. Overall, increasing enzyme exposure time was found to be more detrimental than collagenase concentration for subsequent viability, proliferation, and matrix forming capacity (sGAG and collagen) of these cells resulting in nonuniform cartilaginous matrix deposition. Taken together, consolidating a 3000 U/ml collagenase digest of 1 h at a ratio of 10 ml/g of cartilage tissue with physical agitation cycles can improve efficiency of chondrocyte isolation, yielding robust, more uniform matrix formation.

show abstract

Section: Discussionmentioning

confidence: 99%

Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

Vedicherla

Buckley

2017

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…Various incubation times are used to allow for tissue digestion, but in our experience, longer enzyme exposure times of tissues (and with longer exposures, an increasing number of cells as well) often lead to an increased number of dead cells and/or a lower yield of the cells with a desired phenotype. Considering all mentioned, we used a Trypsin/EDTA combination and an incubation time of 3 h. Although this is not the first research study reporting the use of trypsin for chondrocyte isolation (Hidvegi et al, 2006; Jakob et al, 2001), to the best of our knowledge it is the only one to use only this enzyme during the isolation protocol. Also, the reported incubation time is different to the mentioned studies.…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Naranda

Gradišnik

Gorenjak

et al. 2017

PeerJ

View full text Add to dashboard Cite

BackgroundCartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue.MethodsIsolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production.ResultsCartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well.DiscussionIn this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.

show abstract

“…Each laboratory has a unique method for isolating chondrocytes, involving a number of different enzymes, incubation lengths, and temperatures [17]. Cartilage tissue engineering is moving steadily toward widespread use in vivo; a number of constructs are implanted into humans, and chondrocytes are used in the therapeutic treatment of osteoarthritis.…”

Section: Discussionmentioning

confidence: 99%

Optimization of chondrocyte isolation and characterization for large-scale cartilage tissue engineering

Oseni

Butler

Seifalian

2013

Journal of Surgical Research

View full text Add to dashboard Cite

show abstract

A low temperature method of isolating normal human articular chondrocytes

Cited by 25 publications

References 14 publications

Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Optimization of chondrocyte isolation and characterization for large-scale cartilage tissue engineering

Contact Info

Product

Resources

About