Chondrogenesis of human adipose derived stem cells for future microtia repair using co-culture technique

Goh, Bee See; Omar, Siti Nurhadis Che; Ubaidah, Muhammad Azhan; Saim, Lokman; Sulaiman, Shamsul; Chua, Kien Hui

doi:10.1080/00016489.2016.1257151

Cited by 19 publications

(14 citation statements)

References 20 publications

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“…The question must therefore be asked whether the matrix was articular to begin with. Indeed, in the present study similar gene expression results with respect to COL2A1, SOX9 and ACAN were produced in hADSCs too, supporting the concept that TGF-β isoforms, especially hTGF-β 3 , can induce a form of chondrogenic differentiation also in these stem cells [92]. However, the exception of COL1A1 at day 28 in both pelleted hADSCs in the absence of standard chondrogenic medium (Fig.…”

Section: Discussionsupporting

confidence: 88%

“…This was again substantiated in the present study where 10 ng/ml of hTGF-β 3 promoted significant chondrocyte differentiation and resulted in increased type 2 collagen transcription and translation [16] but in hADSCs. Indeed, most of the previous studies on the articular cartilage differentiation potential were based on MSCs, and were solely focused on whether TGF-β isoforms also possess similar chondrogenic differentiation capacitates as ADSCs [92]. Whilst in vitro traits of TGF-β isoforms at inducing articular chondrogenesis show increased COL2A1, SOX9, ACAN and reduced COL1A1 expressions [93,94], in vivo based research remains problematic; despite extensive investigations demonstrating the potential of MSCs to regenerate cartilage, the latter degenerates quickly in vivo after a certain number of weeks, leading to ossification rather than maintaining articular cartilage state [60,94].…”

Section: Discussionmentioning

confidence: 99%

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Synergistic interaction of hTGF-β3 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

Huang

Seitz²,

Yan

et al. 2020

BMC Biotechnol

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Background Human TGF-β3 has been used in many studies to induce genes coding for typical cartilage matrix components and accelerate chondrogenic differentiation, making it the standard constituent in most cultivation media used for the assessment of chondrogenesis associated with various stem cell types on carrier matrices. However, in vivo data suggests that TGF-β3 and its other isoforms also induce endochondral and intramembranous osteogenesis in non-primate species to other mammals. Based on previously demonstrated improved articular cartilage induction by a using hTGF-β3 and hBMP-6 together on hADSC cultures and the interaction of TGF- β with matrix in vivo, the present study investigates the interaction of a chitosan scaffold as polyanionic polysaccharide with both growth factors. The study analyzes the difference between chondrogenic differentiation that leads to stable hyaline cartilage and the endochondral ossification route that ends in hypertrophy by extending the usual panel of investigated gene expression and stringent employment of quantitative PCR. Results By assessing the viability, proliferation, matrix formation and gene expression patterns it is shown that hTGF-β3 + hBMP-6 promotes improved hyaline articular cartilage formation in a chitosan scaffold in which ACAN with Col2A1 and not Col1A1 nor Col10A1 where highly expressed both at a transcriptional and translational level. Inversely, hTGF-β3 alone tended towards endochondral bone formation showing according protein and gene expression patterns. Conclusion These findings demonstrate that clinical therapies should consider using hTGF-β3 + hBMP-6 in articular cartilage regeneration therapies as the synergistic interaction of these morphogens seems to ensure and maintain proper hyaline articular cartilage matrix formation counteracting degeneration to fibrous tissue or ossification. These effects are produced by interaction of the growth factors with the polysaccharide matrix.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Synergistic interaction of hTGF-β3 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

Huang

Seitz²,

Yan

et al. 2020

BMC Biotechnol

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“…Although other studies have utilized ASC/chondrocyte co-culture for CTE 39 , this report is the first to our knowledge describing successful use of ASC co-culture on 3D-printed tissue engineering scaffolds for successful CTE. The use of ASCs with a co-culture technique is particularly advantageous for a clinically-translatable approach, given the low morbidity to harvest these cells vis-à-vis to bone-marrow derived stromal cells.…”

Section: Discussionmentioning

confidence: 95%

Co‐culture of adipose‐derived stem cells and chondrocytes on three‐dimensionally printed bioscaffolds for craniofacial cartilage engineering

Morrison

Nasser

Kashlan³

et al. 2018

The Laryngoscope

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Objectives/Hypothesis Reconstruction of craniofacial cartilagenous defects are among the most challenging surgical procedures in facial plastic surgery. Bioengineered craniofacial cartilage holds immense potential to surpass current reconstructive options but limitations to clinical translation exist. We endeavored to determine the viability of utilizing adipose-derived stem cell-chondrocyte co-culture and three-dimensional (3D) printing to produce 3D bioscaffolds for cartilage tissue engineering. We describe a feasibility study revealing a novel approach for cartilage tissue engineering with in vitro and in vivo animal data. Methods Porcine adipose-derived stem cells and chondrocytes were isolated and co-seeded at 1:1, 2:1, 5:1, 10:1, and 0:1 experimental ratios in a hyaluronic acid/collagen hydrogel in the pores of 3D-printed polycaprolactone scaffolds to form 3D bioscaffolds for cartilage tissue engineering. Bioscaffolds were cultured in vitro without growth factors for 4 weeks then implanted into the subcutaneous tissue of athymic rats for an additional 4 weeks before sacrifice. Bioscaffolds were subjected to histologic, immunohistochemical, and biochemical analysis. Results Successful production of cartilage was achieved using a co-culture model of adipose-derived stem cells and chondrocytes, without the use of exogenous growth factors. Histology demonstrated cartilage growth for all experimental ratios at the post-in vivo time point confirmed with type II collagen immunohistochemistry. There was no difference in sulfated-glycosaminoglycan production between experimental groups. Conclusion Tissue engineered cartilage was successfully produced on 3D-printed bioresorbable scaffolds using an adipose-derived stem cell and chondrocyte co-culture technique. This potentiates co-culture as a solution for several key barriers to a clinically-translatable cartilage tissue engineering process.

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“…The primer sequences for collagen type II, elastin, aggrecan core protein, and sox9 were listed in Table 1. Primers were chosen from GeneBank data base sequences corresponding to the specific gene Accession Number, according to previously described (Goh et al 2017).…”

Section: Quantitative Gene Expression By Real-time Pcrmentioning

confidence: 99%

Transforming Growth Factor Beta 3 Induced Human Adipose-derived Stem Cells for Auricular Chondrogenesis

Omar

Goh²,

Ubaidah

et al. 2018

JSM

Self Cite

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The limitation of self-repair and proliferation capacity of chondrocytes in cartilage reconstruction lead to alternative search of cell source that can improve the auricular regeneration. Human adipose-derived stem cells (HADSC) are an alternative cell source that have unique characteristics to self-renew and differentiate into various tissues making it suitable for cell therapy and tissue engineering. This study aimed to examine the chondrogenic differentiation potential of (HADSC) in monolayer culture by the presence of different transforming growth factor beta's, TFG-β1,-β2 and-β3. HADSC at passage 3 (1.5 × 10 5 cell/mL) were cultured in chondrogenic medium containing 5 ng/mL of different transforming growth factor beta's, TFG-β1,-β2 and-β3 for 7, 14 and 21 days. Data analysis was evaluated based on the growth rate of cells, cells morphological changed, production of collagen type II and glycosaminoglycan sulphate (sGAG). The quantitative RT-PCR was carried out to determine the chondrogenic, fibrogenic and hypertrophic gene expression levels. Differentiation of HADSC into chondrocytes using TFG-β indicates the occurrence of the chondrogenesis process. The best chondrogenic differentiation was observed in HADSC induced by TFG-β3 through the chondrocytes-like cells morphology with cells aggregation and high production of proteoglycan matrices compared to other TGF-βs groups. Additionally, the expression of chondrocytes-specific genes such as Type II collagen, Aggrecan core protein, Elastin and Sox 9 was high. In conclusion, this study has showed that TGF-β3 is the potential growth factor in producing chondrogenic cells for auricular cartilage tissue engineering.

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Chondrogenesis of human adipose derived stem cells for future microtia repair using co-culture technique

Cited by 19 publications

References 20 publications

Synergistic interaction of hTGF-β3 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

Synergistic interaction of hTGF-β3 with hBMP-6 promotes articular cartilage formation in chitosan scaffolds with hADSCs: implications for regenerative medicine

Co‐culture of adipose‐derived stem cells and chondrocytes on three‐dimensionally printed bioscaffolds for craniofacial cartilage engineering

Transforming Growth Factor Beta 3 Induced Human Adipose-derived Stem Cells for Auricular Chondrogenesis

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