Development of a new co-culture system, the “separable-close co-culture system,” to enhance stem-cell-to-chondrocyte differentiation

Morita, Yasuyuki; Yamamoto, Shuhei; Ju, Yang

doi:10.1007/s10529-015-1858-5

Cited by 9 publications

(6 citation statements)

References 19 publications

(20 reference statements)

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“…Co‐culture systems have received much attention because in the co‐culture techniques, the stem cells that one wants to differentiate are cultured together with target cells in an effort to promote differentiation due to cellular interactions, such as by trophic effects . Recent literature reported by Kraus et al ., indicated that in direct co‐culture, tenascin C was up‐regulated compared with ADMSCs monoculture, and SCX expression was significantly up‐regulated (1.5‐fold) in the group with an ADMSCs‐to‐tenocyte ratio of 1:3.…”

Section: Discussionmentioning

confidence: 99%

Hypoxia enhances tenocyte differentiation of adipose‐derived mesenchymal stem cells by inducing hypoxia‐inducible factor‐1α in a co‐culture system

Zhou

Tien

et al. 2016

Cell Proliferation

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Section: Discussionmentioning

confidence: 99%

Hypoxia enhances tenocyte differentiation of adipose‐derived mesenchymal stem cells by inducing hypoxia‐inducible factor‐1α in a co‐culture system

Zhou

Tien

et al. 2016

Cell Proliferation

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“…Cocultures of mesenchymal stem cells and CHDR can result in improved chondrogenic differentiation in pellets [ 9 ] and immunological interactions including anti-inflammatory regulations attributed to mesenchymal stem cells [ 15 ]. The possible high impact of coculture systems on cartilage tissue engineering is documented by the increasing number of described technical solutions [ 16 ] and experimental designs [ 17 ]. In contrast to previous studies, this investigation focused on an experimental trans-well design using human synovial stem cells.…”

Section: Discussionmentioning

confidence: 99%

The trans-well coculture of human synovial mesenchymal stem cells with chondrocytes leads to self-organization, chondrogenic differentiation, and secretion of TGFβ

et al. 2016

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BackgroundSynovial mesenchymal stem cells (SMSC) possess a high chondrogenic differentiation potential, which possibly supports natural and surgically induced healing of cartilage lesions. We hypothesized enhanced chondrogenesis of SMSC caused by the vicinity of chondrocytes (CHDR).MethodsHuman SMSC and CHDR interactions were investigated in an in-vitro trans-well monolayer coculture over a time period of up to 21 days. Protein expression was analyzed using histology, immunostaining, or enzyme-linked immunosorbent assay. Additionally, mRNA expression was assessed by quantitative PCR.ResultsAfter 7 days, phase-contrast microscopy revealed cell aggregation of SMSC in coculture with CHDR. Afterwards, cells formed spheres and lost adherence. However, this phenomenon was not observed when culturing SMSC alone. Fluorescence labeling showed concurrent collagen type II expression. Addition of transforming growth factor beta (TGFβ) to the cocultures induced SMSC aggregation in less time and with higher intensity. Additionally, alcian blue staining demonstrated enhanced glycosaminoglycan expression around SMSC aggregates after 1 and 2 weeks. Although TGFβ mRNA was expressed in all SMSC, the protein was measured with constantly increasing levels over 21 days only in supernatants of the cocultures. Considering the enhanced mRNA levels following supplementation with TGFβ, a positive feedback mechanism can be supposed. In line with the development of a chondrogenic phenotype, aggrecan mRNA expression increased after 7 and 14 days in the cocultures with and without TGFβ. Coculture conditions also amplified collagen type II mRNA expression after 2 weeks without and already after 1 week with TGFβ. There was no difference in collagen type I and type X expression between SMSC alone and the coculture with CHDR. Expression of both collagens increased following addition of TGFβ. mRNA data correlated with the intensity of immunofluorescence staining.ConclusionsParacrine effects of CHDR induce a chondrogenic phenotype in SMSC possibly mimicking joint homeostasis. Coculture approaches may lead to a better understanding of cellular interactions with potential implications for cartilage repair procedures.

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“…There are currently several co-culture models being usedincluding mixed coculture, separated co-culture, and bothto investigate the mechanism of co-culture capable of supporting stable chondrogenesis (Levorson et al 2014;Morita et al 2015). However, overall, the mechanism is still under debate.…”

Section: Co-culture Of Chondrocytes With Mscsmentioning

confidence: 99%

Generation of Ear Cartilage for Auricular Reconstruction

Liu¹,

Cao²

2020

Organ Tissue Engineering

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Auricular reconstruction is among the most challenging surgical procedures in plastic and reconstructive surgery because of the lack of an ideal auricle substitute that can guarantee a long-lasting outcome while involving minimal donor site morbidity. Tissue-engineered cartilage may provide an ideal autologous solution. After the first report on generation of human ear-shaped cartilage in a nude mouse model in 1997, cartilages with human ear shape have been engineered in vitro, in nude mice, and in immunocompetent animals using various cells and scaffolds. Recently, our group reported a pilot clinical trial of in vitro engineered human earshaped cartilage and its clinical translation for auricular reconstruction. This bench-to-bed process spanning over the past two decades can provide an indepth understanding of the development of cartilage tissue engineering for

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Development of a new co-culture system, the “separable-close co-culture system,” to enhance stem-cell-to-chondrocyte differentiation

Abstract: The new co-culture system is effective in terms of assessing gene expression in two cell types.

Cited by 9 publications

References 19 publications

Hypoxia enhances tenocyte differentiation of adipose‐derived mesenchymal stem cells by inducing hypoxia‐inducible factor‐1α in a co‐culture system

Hypoxia enhances tenocyte differentiation of adipose‐derived mesenchymal stem cells by inducing hypoxia‐inducible factor‐1α in a co‐culture system

The trans-well coculture of human synovial mesenchymal stem cells with chondrocytes leads to self-organization, chondrogenic differentiation, and secretion of TGFβ

Generation of Ear Cartilage for Auricular Reconstruction

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