The effect of temporal manipulation of transforming growth factor beta 3 and fibroblast growth factor 2 on the derivation of proliferative chondrocytes from mensenchymal stem cells—A study monitored by quantitative reverse transcription polymerase chain reaction and molecular beacon based nanosensors

Tay, Li Min; Wiraja, Christian; Wu, Yingnan; Yang, Zheng; Lee, Eng Hin; Xu, Chen

doi:10.1002/jbm.a.36286

Cited by 8 publications

(8 citation statements)

References 35 publications

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“…For chondrocyte taxonomy, some studies have shown chondrocyte subtypes mainly included ProCs, preHTCs and HTCs [29][30][31]. In addition to these three common chondrocyte subtypes, QB and his partners rst identi ed 4 new populations of chondrocytes in the human OA cartilage by performing unbiased transcriptome-wide scRNA-seq analysis and computational analysis on some chondrocytes from 10 OA patients [16].…”

Section: Discussionmentioning

confidence: 99%

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Zhang

Huang

et al. 2020

Preprint

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Background： Single-cell RNA sequencing (scRNA-seq) was recently adopted for exploring molecular programmes and lineage progression patterns of pathogenesis of important diseases. In this study, scRNA-seq was used to identify potential markers for chondrocytes in osteoarthritis (OA) and to explore the function of different types of chondrocytes in OA.Methods：Here we aimed to identify the biomarkers and differentiation of chondrocyte by Single-cell RNA seq analysis. GeneOntology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to identify the function of candidate marker genes in chondrocytes. Protein–protein interaction (PPI) network was constructed to find the hub genes in 3 types of chondrocyte respectively. We also used qRT-PCR to detect the expression level of the candidate marker genes in different types of chondrocyte. Results： In this study, we characterized the single-cell expression profiling of 480 chondrocyte samples and found hypertrophic chondrocyte (HTC), homeostatic chondrocyte (HomC) and fibrocartilage chondrocyte (FC) respectively. The results of GO and KEGG analysis showed the candidate marker genes made specific function in these chondrocytes to regulate the development of OAs respectively. We further revealed the differential expression of top 10 marker genes in 3 types of chondrocyte. The marker genes of HTC and FC were mainly expressed in their cell subset respectively. The marker genes of HomC did not have obviously differential expression among different types of chondrocyte. Last, we predicted the key genes in each cell subset. CD44, JUN and FN1 were predicted tightly related to the proliferation and differentiation of chondrocytes in OAs and could be regarded as biomarkers to estimate the development of OA. Conclusion： Our results provide new insights into exploring the roles of different types of chondrocyte in OA. The biomarkers of chondrocyte were also valuable for estimating OA progression.

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Section: Discussionmentioning

confidence: 99%

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Zhang

Huang

et al. 2020

Preprint

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“…For chondrocyte taxonomy, some studies have shown chondrocyte subtypes mainly included ProCs, preHTCs and HTCs [29][30][31]. In addition to these three common chondrocyte subtypes, Ji et al rst identi ed 4 new populations of chondrocytes in the human OA cartilage by performing unbiased transcriptome-wide scRNA-seq analysis and computational analysis on some chondrocytes from 10 OA patients [16].…”

Section: Discussionmentioning

confidence: 99%

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Zhang

Huang

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Single-cell RNA sequencing (scRNA-seq) was recently adopted for exploring molecular programmes and lineage progression patterns of pathogenesis of important diseases. In this study, scRNA-seq was used to identify potential markers for chondrocytes in osteoarthritis (OA) and to explore the function of different types of chondrocytes in OA. Methods: Here we aimed to identify the biomarkers and differentiation of chondrocyte by Single-cell RNA seq analysis. GeneOntology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to identify the function of candidate marker genes in chondrocytes. Protein-protein interaction (PPI) network was constructed to find the hub genes in 3 types of chondrocyte respectively. We also used qRT-PCR to detect the expression level of the candidate marker genes in different types of chondrocyte. Results: In this study, we characterized the single-cell expression profiling of 480 chondrocyte samples and found hypertrophic chondrocyte (HTC), homeostatic chondrocyte (HomC) and fibrocartilage chondrocyte (FC) respectively. The results of GO and KEGG analysis showed the candidate marker genes made specific function in these chondrocytes to regulate the development of OAs respectively. We further revealed the differential expression of top 10 marker genes in 3 types of chondrocyte. The marker genes of HTC and FC were mainly expressed in their cell subset respectively. The marker genes of HomC did not have obviously differential expression among different types of chondrocyte. Last, we predicted the key genes in each cell subset. CD44, JUN and FN1 were predicted tightly related to the proliferation and differentiation of chondrocytes in OAs and could be regarded as biomarkers to estimate the development of OA. Conclusion: Our results provide new insights into exploring the roles of different types of chondrocyte in OA. The biomarkers of chondrocyte were also valuable for estimating OA progression.

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“…The concurrent application of hypoxia and FGF-2 could provide a favorable condition for culturing human bone marrow-derived stem cells to be used in clinical applications associated with bone tissue engineering, due to the enhancement of cellular proliferation and regenerative potential (24). The effects of temporal manipulation were achieved by applying FGF-2 and transforming growth factor-β3 on the derivation of proliferative chondrocytes from mesenchymal stem cells (25).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

Tae

Park

2019

Exp Ther Med

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Fibroblast growth factor-2 (FGF-2) is reported to have various functions and is considered a key human mesenchymal stem cell mitogen, often supplemented to increase human mesenchymal stem cell growth rates. The purpose of this study was to evaluate the effects of FGF-2 on cellular viability and osteogenic differentiation using three-dimensional cell spheroids of stem cells. Three-dimensional cell spheroids were fabricated using concave silicon elastomer-based microwells in the presence of FGF-2 at concentrations of 0, 30, 60 and 90 ng/ml. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit-8 assay. Alkaline phosphatase activity and Alizarin Red S staining were used to assess osteogenic differentiation. Spheroids were well formed in silicon elastomer-based concave microwells on Day 1. The average spheroid diameters at Day 1 for FGF-2 at 0, 30, 60 and 90 ng/ml were 202.2±3.0, 206.6±22.6, 208.8±6.8 and 196.6±26.7 µm, respectively (P>0.05). The majority of the cells in the cell spheroids emitted green fluorescence. The relative Cell Counting Kit-8 assay values for FGF-2 at 0, 30, 60 and 90 ng/ml at Day 1 were 100.0±5.5, 101.8±8.8, 99.2±4.8 and 103.4±9.6% (P>0.05). The addition of FGF-2 at 60 ng/ml concentration produced the highest value for alkaline phosphatase activity. Mineralized extracellular deposits were evenly observed in each group, and the highest value was identified for FGF-2 groups at 60 ng/ml concentration for Alizarin Red S staining. Based on these findings, it was concluded that FGF-2 may increase alkaline phosphatase activity or Alizarin Red S staining, and further studies are needed to fully elucidate the mechanisms of FGF-2.

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Cited by 8 publications

References 35 publications

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Single-cell RNA seq analysis identifies the biomarkers and differentiation of chondrocyte in human osteoarthritis

Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

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