Expansion of human nasal chondrocytes on macroporous microcarriers enhances redifferentiation

Malda, Jos; Kreijveld, Ellen; Temenoff, Johnna S.; Blitterswijk, Clemens A. van; Riesle, Jens

doi:10.1016/s0142-9612(03)00428-9

Cited by 127 publications

(108 citation statements)

References 41 publications

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“…6 Cells were seeded at a density of 10 4 cells per mg microcarrier per ml culture medium and cultivated using the Variomag BIOSYSTEM 4B (Thermo Fisher, Waltham, MA) under intermittent stirring at 20 rpm for 30 s every 30 min during the first 2 days. The stirring regime was then changed to continuous stirring at 50-65 rpm for the remainder of culture.…”

Section: Cell Expansionmentioning

confidence: 99%

“…Microcarrier cultures were supplemented with 1 mCi/ml L- Cell Viability, Metabolic Activity, and Actin Filaments To characterize cell viability in microcarrier cultures, cell/ microcarrier samples were gently washed twice in PBS, incubated for 5 min at 378C in PBS containing 0.67 mg/ml fluorescein diacetate (FDA) and 5 mg/ml propidium iodide (PI) (both Invitrogen) and washed again in PBS. 6 Metabolically active cells were visualized by using a 1 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution. 6 Phase contrast and epi-fluorescence images were captured by a digital camera mounted on an Eclipse TS100 microscope with T-FL epi-fluorescence attachment (Nikon, Tokyo, Japan).…”

Section: Chondrogenic Differentiationmentioning

confidence: 99%

“…6 Metabolically active cells were visualized by using a 1 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution. 6 Phase contrast and epi-fluorescence images were captured by a digital camera mounted on an Eclipse TS100 microscope with T-FL epi-fluorescence attachment (Nikon, Tokyo, Japan).…”

Section: Chondrogenic Differentiationmentioning

confidence: 99%

“…4,5 The use of microcarriers for chondrocyte expansion has been investigated to overcome some of these effects. [6][7][8][9][10] Microcarrier culture offers a cost-efficient method of propagating large populations of cells due to a high surface area to volume ratio. 10 In addition, microcarriers may be cultured in bioreactors which allow critical biological and chemical factors (pH, pO 2 , and metabolites), as well as physical factors (such as shear), to be monitored and precisely controlled.…”

mentioning

confidence: 99%

“…In particular, porous collagen-derived microcarriers have been shown to benefit the expansion of bovine articular chondrocytes 7,8 and enhance the chondrogenic potential of human nasoseptal chondrocytes. 6 However, these cells are dissimilar to human articular chondrocytes, [11][12][13] thus it remains unclear if expansion on gelatin microcarriers improves the chondrogenic potential of clinically relevant adult human articular chondrocytes.…”

mentioning

confidence: 99%

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Adult human articular chondrocytes in a microcarrier‐based culture system: expansion and redifferentiation

Schrobback

Klein

Schuetz

et al. 2010

Journal Orthopaedic Research

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Expanding human chondrocytes in vitro while maintaining their ability to form cartilage remains a key challenge in cartilage tissue engineering. One promising approach to address this is to use microcarriers as substrates for chondrocyte expansion. While microcarriers have shown beneficial effects for expansion of animal and ectopic human chondrocytes, their utility has not been determined for freshly isolated adult human articular chondrocytes. Thus, we investigated the proliferation and subsequent chondrogenic differentiation of these clinically relevant cells on porous gelatin microcarriers and compared them to those expanded using traditional monolayers. Chondrocytes attached to microcarriers within 2 days and remained viable over 4 weeks of culture in spinner flasks. Cells on microcarriers exhibited a spread morphology and initially proliferated faster than cells in monolayer culture, however, with prolonged expansion they were less proliferative. Cells expanded for 1 month and enzymatically released from microcarriers formed cartilaginous tissue in micromass pellet cultures, which was similar to tissue formed by monolayer-expanded cells. Cells left attached to microcarriers did not exhibit chondrogenic capacity. Culture conditions, such as microcarrier material, oxygen tension, and mechanical stimulation require further investigation to facilitate the efficient expansion of clinically relevant human articular chondrocytes that maintain chondrogenic potential for cartilage regeneration applications. ß

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Section: Cell Expansionmentioning

confidence: 99%

Section: Chondrogenic Differentiationmentioning

confidence: 99%

Section: Chondrogenic Differentiationmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Adult human articular chondrocytes in a microcarrier‐based culture system: expansion and redifferentiation

Schrobback

Klein

Schuetz

et al. 2010

Journal Orthopaedic Research

View full text Add to dashboard Cite

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Biodegradable Polymers for Emerging Clinical Use in Tissue Engineering

Verma

Garkhal

Mittal

et al. 2011

Biodegradable Polymers in Clinical Use and Clinical Development

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Silk‐inspired polyurethane containing GlyAlaGlyAla tetrapeptide. I. Synthesis and primary structure

Liu

Zhao

et al. 2010

J of Applied Polymer Sci

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Biomedical polyurethane (PU) and silk fibroin have similar molecular architecture in their primary and aggregate structure, both of which have imido bonds and microphase separation, and they have been employed as scaffold materials for biomedical applications. As the featured peptide sequence of silkworm silk protein, GlyAlaGlyAla (GAGA) tetrapeptide was synthesized by using traditional liquid-phase peptide synthesis method with Boc-protected alanine and glycine as starting materials, and was transformed to its derivative with two end-amido groups. The derivative was incorporated into the backbone of the classical synthetic copolyurethane as a chain extender to form a silk-inspired polyurethane. Elemental analysis showed the nitrogen content in the silk-inspired PU was higher than that in the butanediol-extended PU but lower than the theoretical value, indicating the chain extender containing GAGA was partially incorporated into the PU chain. The results from 1 H-nuclear magnetic resonance, Fourier transform infrared spectra, Raman scattering spectra and pyrolysis-gas chromatography/mass spectrometry qualitatively identified the primary structure of the silk-inspired polyurethane. This concept and strategy may allow the fabrication of a new class of thermoplastic polyurethane elastomer to mimic the structure and properties of silk fibers if in fiber form, which combines traditional synthetic polymer chains with peptide sequences.

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Expansion of human nasal chondrocytes on macroporous microcarriers enhances redifferentiation

Cited by 127 publications

References 41 publications

Adult human articular chondrocytes in a microcarrier‐based culture system: expansion and redifferentiation

Adult human articular chondrocytes in a microcarrier‐based culture system: expansion and redifferentiation

Biodegradable Polymers for Emerging Clinical Use in Tissue Engineering

Silk‐inspired polyurethane containing GlyAlaGlyAla tetrapeptide. I. Synthesis and primary structure

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