Enhanced proliferation and differentiation of human articular chondrocytes when seeded at low cell densities in alginate <i>In Vitro</i>

Gagne, T. A.; Chappell-Afonso, K.; Johnson, JL; McPherson, John M.; Oldham, Carla; Tubo, Ross; Vaccaro, Carlos; Vasios, George

doi:10.1002/jor.1100180606

Cited by 45 publications

(58 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The differential expression of the cytokine CTGF by adult articular chondrocytes, within 48 hr of transfer to suspension culture at 10 6 cells/ml ( Fig. 2A) was consistent with relatively low level of cell proliferation observed in suspension culture (Gagne et al, 2000). CTGF expression had been previously identified in chondrocytes derived from growth plate and a human chondrosarcoma, but not in adult articular chondrocytes (Nakanishi et al, 1997).…”

Section: Discussionsupporting

confidence: 82%

See 1 more Smart Citation

Differential expression of multiple genes during articular chondrocyte redifferentiation

Haudenschild¹,

McPherson²,

Tubo³

et al. 2001

Anat. Rec.

View full text Add to dashboard Cite

Articular chondrocytes undergo a rapid change in phenotype and gene expression, termed dedifferentiation, when isolated from cartilage tissue and cultured on tissue culture plastic. On the other hand, "redifferentiation" of articular chondrocytes in suspension culture is characterized by decreased cellular proliferation and the reinitiation of synthesis of hyaline articular cartilage extracellular matrix molecules. The molecular triggers for these events have yet to be defined. Subtracted cDNA libraries representing genes involved in the early events of adult human articular chondrocyte redifferentiation were generated from human articular chondrocytes that were first cultured in monolayer, and subsequently transferred to suspension culture at 10 6 cells/ml for redifferentiation. Differential regulation of genes involved in cellular organization, nuclear structure, cellular growth regulation, and extracellular matrix deposition and remodeling were observed within 48 hr of this transfer. Many of these genes had not been previously identified in the chondrocyte differentiation pathway and a number of the isolated cDNAs did not have homologies to sequences in the public data banks. Genes involved in IL-6 signal transduction including acute phase response factor (APRF), Mn superoxide dismutase, and IL-6 itself were up-regulated in suspension culture. Membrane glycoprotein gp130, a component of the IL-6 receptor, was down-regulated. Other genes involved in cell polarity, cell adherence, apoptosis, and possibly TGF-beta signaling were differentially regulated. The differential regulation of the cytokine connective tissue growth factor (CTGF) during the early stages of articular chondrocyte redifferentiation, decreasing within 48 hours of transfer to suspension culture, was particularly interesting given its reported role in the stimulation of cellular proliferation. CTGF was highly expressed in proliferative monolayer culture, and then greatly reduced by redifferentiation in standard high-density suspension culture. When articular chondrocytes were seeded in suspension at low-density (10 4 cells/ml), however, high levels of CTGF were observed along with increased levels of mature articular cartilage extracellular matrix protein RNAs, such as type II collagen and aggrecan. Although the role of CTGF in articular cartilage biology remains to be elucidated, the results described here demonstrate the potential utility of subtractive hybridization in understanding the process of articular chondrocyte redifferentiation. Anat Rec 263: [91][92][93][94][95][96][97][98] 2001.

show abstract

Section: Discussionsupporting

confidence: 82%

“…osition (Gagne et al, 2000). Northern blot analysis of mRNA from articular chondrocyte suspension cultures initially seeded at 10 4 , 10 5 , and 10 6 cell/ml revealed that the level of CTGF expressed was higher in suspension cultures seeded at lower cell density (Fig.…”

Section: Discussionmentioning

confidence: 92%

Differential expression of multiple genes during articular chondrocyte redifferentiation

Haudenschild¹,

McPherson²,

Tubo³

et al. 2001

Anat. Rec.

View full text Add to dashboard Cite

show abstract

“…This result could be related to the post-mortem time when autopsy tissues were processed (within 2 4 4 8 h) whilst DAC was processed within 12 h after arthroscopic retrieval from patients during an ACI procedure. Articular cartilage from autopsies should be processed for cell culture within 24 h post-mortem [28] or within 48 h [9]. Although it has been shown that 88-96'%1 post-mortem human chondrocytes, kept at 4 "C within the first 4.5 days, are viable as observed by vital staining and confocal laser scanning 3D microscopy [16].…”

Section: Discussionmentioning

confidence: 99%

The use of debrided human articular cartilage for autologous chondrocyte implantation: Maintenance of chondrocyte differentiation and proliferation in type I collagen gels

Chaipinyo

Oakes

Damme

2004

Journal Orthopaedic Research

View full text Add to dashboard Cite

Autologous chondrocyte implantation (ACI) is the most promising surgical treatment for large full thickness knee joint articular cartilage (AC) defects where cells from healthy non-weight bearing area AC are multiplied in vitro and implanted into such defects. In the routine surgical procedure for symptomatic knee full thickness AC defects, damaged AC surrounding the edge and the base of such defects is usually debrided and discarded. The purpose of this study was to examine if chondrocytes from this 'debrided' AC can proliferate, synthesize a cartilage specific matrix and thus can be used for ACI. Methods: Biopsies were retrieved from 12 patients (debrided articular cartilage: DAC, aged and from two autopsies (normal articular cartilage: NAC, aged 21 and 25). Chondrocytes were isolated, seeded at low density in type I collagen gels and as monolayer cultures for 4 weeks without passage.Results: After 4 weeks cultures in type I collagen gels, cell proliferation from DAC (18.34k 1.95 fold) was similar to cells from NAC ( 1 1.24 f 1.02 fold). Syntheses of proteoglycan and collagen in DAC were also similar to NAC. Newly synthesized matrices in gel cultures consisted predominantly of type I1 collagen as shown by immuno-labelling and SDS-PAGE followed by fluorography. Chondrocytes from 'debrided human AC' cultured at low density in type I collagen gels may be used for the ACI procedure as they provide sufficient viable cell numbers for ACI and maintain their chondrocyte phenotype as they synthesize a cartilage-like matrix.

show abstract

“…The heterogeneity of articular chondrocytes may also play a role in cell survival, proliferation and matrix synthesis [Panasyuk et al, 2004]. In addition, mitotic activity, seeded cell density and ECM composition may play important roles inasmuch as mutual infl uences are not fully understood yet [Paige et al, 1995;Gagne et al, 2000;Qi and Scully, 2003].…”

Section: Hydrogel Culturementioning

confidence: 99%

Morphology and Function of Ovine Articular Cartilage Chondrocytes in 3-D Hydrogel Culture

Schagemann

Mrosek

Landers

et al. 2006

Cells Tissues Organs

View full text Add to dashboard Cite

Different cell- and biomaterial-based tissue engineering techniques are under investigation to restore damaged tissue. Strategies that use chondrogenic cells or tissues in combination with bioresorbable delivery materials are considered to be suitable to regenerate bio-artificial cartilage. Three-dimensional (3-D) cell embedding techniques can provide anchorage-independent cell growth and homogenous spatial cell arrangement, which play a key role in the maintenance of the characteristic phenotype and thus the formation of differentiated tissue. We developed a new injectable high water content (90%) hydrogel formulation with 5% sodium alginic acid and 5% gelatin as a temporary supportive intercellular matrix for 3-D cell culture. The objective was to determine whether the in vitro hydrogel culture of chondrocytes could preserve hyaline characteristics and thus could provide cartilage regeneration in vitro. Chondrocytes harvested from knee joints of skeletally mature sheep were cultured 3-D in hydrogel (7 × 10⁶ cells/ml, 2.8-µl beads) for up to 10 weeks. Cell morphology and viability were evaluated with light microscopy, and proliferative activity was assessed with antibromodeoxyuridine immunofluorescence. Expression of collagens type I (COL1) and II (COL2), cartilage proteoglycans (PG) and hyaluronan synthases (HAS) were studied immunohistochemically. We observed that up to 36% of chondrocytes proliferated, while almost 100% presented a differentiated spheroidal phenotype. After an initial decrease at 2 weeks, cell density recovered to 85% of the initial absolute value at 10 weeks. Expression of hyaline matrix molecules resembled the in vivo pattern with increasing spatial deposition of PG and COL2. The proportion of PG-positive cells increased from initially 13 to 53% after 10 weeks, in contrast to consistently 100% COL2-positive cells. We conclude that 3-D hydrogel culture, even without mechanical stimulation or growth factor application, can keep chondrocytes in a differentiated state and provides a chondrogenic cell environment for in vitro cartilage regeneration for at least 10 weeks. Moreover, this hydrogel appears to be a suitable cell delivery material for subsequent in vivo implantation.

show abstract

Enhanced proliferation and differentiation of human articular chondrocytes when seeded at low cell densities in alginate In Vitro

Cited by 45 publications

References 28 publications

Differential expression of multiple genes during articular chondrocyte redifferentiation

Differential expression of multiple genes during articular chondrocyte redifferentiation

The use of debrided human articular cartilage for autologous chondrocyte implantation: Maintenance of chondrocyte differentiation and proliferation in type I collagen gels

Morphology and Function of Ovine Articular Cartilage Chondrocytes in 3-D Hydrogel Culture

Contact Info

Product

Resources

About