Proliferating Cells versus Differentiated Cells in Tissue Engineering

Strehl, Raimund; Schumacher, Karl; Vries, Uwe de; Minuth, Will W.

doi:10.1089/107632702753503036

Cited by 58 publications

(48 citation statements)

References 27 publications

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“…The present data might recite the fact that while chondrocytes preferentially expanded in monolayer, they rather stayed unchanged in number under 3D culture conditions. 28 Based on the decrease in DNA content in the IHE group at 4 weeks, we consider that the seeded chondrocytes were unstably attached on the epithelium of IHE (Fig. 3A and D); thus the cells might be lost by shearing force generated by change of media during long-term culture.…”

Section: Discussionmentioning

confidence: 99%

Human Amniotic Membrane as a Delivery Matrix for Articular Cartilage Repair

Jin

Park²,

Choi³

et al. 2007

Tissue Engineering

113

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The purpose of this study is to evaluate the feasibility of human amniotic membrane (HAM) as a chondrocyte carrier by assessing cell proliferation and maintenance of phenotype in vitro and cartilage regeneration in vivo. Intact HAM was treated with 0.1% trypsin-ethylenediaminetetraacetic acid (EDTA) for 15 min and the epithelial cells removed to make a denuded HAM. Rabbit articular chondrocytes were then seeded on three different HAM substrates: the epithelial side of intact HAM (IHE), basement side of denuded HAM (DHB), and stromal side of denuded HAM (DHS). These cell-substrate specimens were cultured for up to 4 weeks, and cell proliferation rate and phenotypic stability were examined at weeks 1 and 4. While chondrocytes grew in monolayer fashion on the surface of IHE and DHB substrates, the cells seeded in DHS penetrated and spread into the whole thickness of the stromal layer. The proliferating activity of chondrocytes in DHB was continuously up-regulated. A similar proliferating activity was observed in DHS in the first week, which remained stable for up to 4 weeks. The expression of type II collagen gradually increased with time in the DHS group, while it gradually decreased in the DHB group or was not detected at all in the IHE group. These results suggested that denuded HAM was able to support chondrocyte proliferation and maintenance of phenotype in vitro, seemingly more favorable when DHS was used. Based on this data, the DHS with chondrocytes was used to cover rabbit osteochondral defect with the stromal side facing in. The defect area was successfully regenerated with hyaline cartilage in the Safranin-O stain and International Cartilage Repair Society (ICRS) scoring after 8 weeks of implantation. In conclusion, our findings suggest that denuded HAM could be one of the ideal cell carrier matrices for cartilage regeneration.

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Section: Discussionmentioning

confidence: 99%

Human Amniotic Membrane as a Delivery Matrix for Articular Cartilage Repair

Jin

Park²,

Choi³

et al. 2007

Tissue Engineering

113

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“…This information is highly valuable when producing bone TE constructs by procedures that distinguish between cell proliferation and cell differentiation or that depend on cells reaching a certain critical cell density. 49 The DNA results of the cultured 3D TE constructs were not influenced by the culture condition (static versus dynamic), while the metabolic activity of the static cultured TE constructs was significantly different from the dynamically cultured scaffolds (Fig. 5C).…”

mentioning

confidence: 86%

Noninvasive Real-Time Monitoring by AlamarBlue^®DuringIn VitroCulture of Three-Dimensional Tissue-Engineered Bone Constructs

Zhou

Holsbeeks

Impens

et al. 2013

Tissue Engineering Part C: Methods

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Bone tissue engineering (TE) aims to develop reproducible and predictive three-dimensional (3D) TE constructs, defined as cell-seeded scaffolds produced by a controlled in vitro process, to heal or replace damaged and nonfunctional bone. To control and assure the quality of the bone TE constructs, a prerequisite for regulatory authorization, there is a need to develop noninvasive analysis techniques to evaluate TE constructs and to monitor their behavior in real time during in vitro culturing. Most analysis techniques, however, are limited to destructive end-point analyses. This study investigates the use of the nontoxic alamarBlue Ò (AB) reagent, which is an indicator for metabolic cell activity, for monitoring the cellularity of 3D TE constructs in vitro as part of a bioreactor culturing processes. Within the field of TE, bioreactors have a huge potential in the translation of TE concepts to the clinic. Hence, the use of the AB reagent was evaluated not only in static cultures, but also in dynamic cultures in a perfusion bioreactor setup. Hereto, the AB assay was successfully integrated in the bioreactor-driven TE construct culture process in a noninvasive way. The obtained results indicate a linear correlation between the overall metabolic activity and the total DNA content of a scaffold upon seeding as well as during the initial stages of cell proliferation. This makes the AB reagent a powerful tool to follow-up bone TE constructs in real-time during static as well as dynamic 3D cultures. Hence, the AB reagent can be successfully used to monitor and predict cell confluence in a growing 3D TE construct.

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“…Initially, during the in vitro culture time, proliferation is necessary to achieve a high number of cells. However, when implanted into a host, a quiescent, differentiated and contractile phenotype is preferable [32]. Vascular smooth muscle cells cultured in vitro typically revert from a contractile phenotype to a synthetic phenotype.…”

Section: Article In Pressmentioning

confidence: 99%