scholarly journals Evaluation of Insulin Medium or Chondrogenic Medium on Proliferation and Chondrogenesis of ATDC5 Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Yongchang Yao ◽  
Zhichen Zhai ◽  
Yingjun Wang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Line ◽  
Growth Medium ◽  
Cartilage Repair ◽  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cell Model ◽  
Chondrogenic Medium ◽  
Atdc5 Cell

Background. The ATDC5 cell line is regarded as an excellent cell model for chondrogenesis. In most studies with ATDC5 cells, insulin medium (IM) was used to induce chondrogenesis while chondrogenic medium (CM), which was usually applied in chondrogenesis of mesenchymal stem cells (MSCs), was rarely used for ATDC5 cells. This study was mainly designed to investigate the effect of IM, CM, and growth medium (GM) on chondrogenesis of ATDC5 cells.Methods. ATDC5 cells were, respectively, cultured in IM, CM, and GM for a certain time. Then the proliferation and the chondrogenesis progress of cells in these groups were analyzed.Results. Compared with CM and GM, IM promoted the proliferation of cells significantly. CM was effective for enhancement of cartilage specific markers, while IM induced the cells to express endochondral ossification related genes. Although GAG deposition per cell in CM group was significantly higher than that in IM and GM groups, the total GAG contents in IM group were the most.Conclusion. This study demonstrated that CM focused on induction of chondrogenic differentiation while IM was in favor of promoting proliferation and expression of endochondral ossification related genes. Combinational use of these two media would be more beneficial to bone/cartilage repair.

scholarly journals Articular chondrocyte derived exosomes promote cartilage differentiation of human umbilical cord mesenchymal stem cells by activation of autophagy

2020 ◽  
Author(s):  
Ke Ma ◽  
Bo Zhu ◽  
Zetao Wang ◽  
Peian Cai ◽  
Mingwei He ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Cartilage Repair ◽  
Chondrogenic Differentiation ◽  
Phenotypic Expression ◽  
Cartilage Regeneration ◽  
Human Umbilical Cord ◽  
Articular Chondrocyte

Abstract Background Umbilical cord mesenchymal stem cells (HUCMSCs)-based therapies were previously predicated in cartilage regeneration due to the chondrogenic potential of MSCs. However, chondrogenic differentiation of HUMSCs is limited by administration of growth factors like TGF-β that may cause cartilage hypertrophy. It has been reported the exosomes could modulate phenotypic expression of stem cells. However, the role of human chondrogenic derived exosomes (C-EXO) in chondrogenic differentiation of HUCMSCs has not been reported. Results In this study, we successfully isolated chondrocyte-derived exosomes (C-EXO) from human multi-finger cartilage and found that C-EXO efficiently promoted the proliferation and chondrogenic differentiation of HUCMSCs, evidenced by highly expressed aggrecan (ACAN), COL2A and SOX-9. Also, the expression of the fibrotic marker, COL1A and hypertrophic marker, COL10, was significantly lower than that induced by TGF-β. In vivo, stimulation of C-EXO accelerated HUCMSCs-mediated cartilage repair in rabbit models. Furthermore, C-EXO led to increasing autophagosomes during the process of chondrogenic differentiation, indicating that C-EXO promoted cartilage regeneration might be through the activation of autophagy. Conclusions This study suggests that C-EXO has an essential role in fostering chondrogenic differentiation and proliferation of HUCMSCs, which may be a stable supply for articular cartilage repair.

scholarly journals Inability of Low Oxygen Tension to Induce Chondrogenesis in Human Infrapatellar Fat Pad Mesenchymal Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Samia Rahman ◽  
Alexander R. A. Szojka ◽  
Yan Liang ◽  
Melanie Kunze ◽  
Victoria Goncalves ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Oxygen Tension ◽  
Endochondral Ossification ◽  
Infrapatellar Fat Pad ◽  
Low Oxygen ◽  
Fat Pad ◽  
Chondrogenic Medium ◽  
Low Oxygen Tension

ObjectiveArticular cartilage of the knee joint is avascular, exists under a low oxygen tension microenvironment, and does not self-heal when injured. Human infrapatellar fat pad-sourced mesenchymal stem cells (IFP-MSC) are an arthroscopically accessible source of mesenchymal stem cells (MSC) for the repair of articular cartilage defects. Human IFP-MSC exists physiologically under a low oxygen tension (i.e., 1–5%) microenvironment. Human bone marrow mesenchymal stem cells (BM-MSC) exist physiologically within a similar range of oxygen tension. A low oxygen tension of 2% spontaneously induced chondrogenesis in micromass pellets of human BM-MSC. However, this is yet to be demonstrated in human IFP-MSC or other adipose tissue-sourced MSC. In this study, we explored the potential of low oxygen tension at 2% to drive the in vitro chondrogenesis of IFP-MSC. We hypothesized that 2% O2 will induce stable chondrogenesis in human IFP-MSC without the risk of undergoing endochondral ossification at ectopic sites of implantation.MethodsMicromass pellets of human IFP-MSC were cultured under 2% O2 or 21% O2 (normal atmosphere O2) in the presence or absence of chondrogenic medium with transforming growth factor-β3 (TGFβ3) for 3 weeks. Following in vitro chondrogenesis, the resulting pellets were implanted in immunodeficient athymic nude mice for 3 weeks.ResultsA low oxygen tension of 2% was unable to induce chondrogenesis in human IFP-MSC. In contrast, chondrogenic medium with TGFβ3 induced in vitro chondrogenesis. All pellets were devoid of any evidence of undergoing endochondral ossification after subcutaneous implantation in athymic mice.

scholarly journals HIF-1α as a Regulator of BMP2-Induced Chondrogenic Differentiation, Osteogenic Differentiation, and Endochondral Ossification in Stem Cells

2015 ◽  
Vol 36 (1) ◽  
pp. 44-60 ◽  
Author(s):  
Nian Zhou ◽  
Ning Hu ◽  
Jun-Yi Liao ◽  
Liang-Bo Lin ◽  
Chen Zhao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Cartilage Repair ◽  
Chondrogenic Differentiation ◽  
Endochondral Ossification ◽  
Cartilage Tissue Engineering ◽  
Bone Morphogenetic Protein 2 ◽  
Cartilage Tissue ◽  
Cartilage Formation

Background/Aims: Joint cartilage defects are difficult to treat due to the limited self-repair capacities of cartilage. Cartilage tissue engineering based on stem cells and gene enhancement is a potential alternative for cartilage repair. Bone morphogenetic protein 2 (BMP2) has been shown to induce chondrogenic differentiation in mesenchymal stem cells (MSCs); however, maintaining the phenotypes of MSCs during cartilage repair since differentiation occurs along the endochondral ossification pathway. In this study, hypoxia inducible factor, or (HIF)-1α, was determined to be a regulator of BMP2-induced chondrogenic differentiation, osteogenic differentiation, and endochondral bone formation. Methods: BMP2 was used to induce chondrogenic and osteogenic differentiation in stem cells and fetal limb development. After HIF-1α was added to the inducing system, any changes in the differentiation markers were assessed. Results: HIF-1α was found to potentiate BMP2-induced Sox9 and the expression of chondrogenesis by downstream markers, and inhibit Runx2 and the expression of osteogenesis by downstream markers in vitro. In subcutaneous stem cell implantation studies, HIF-1α was shown to potentiate BMP2-induced cartilage formation and inhibit endochondral ossification during ectopic bone/cartilage formation. In the fetal limb culture, HIF-1α and BMP2 synergistically promoted the expansion of the proliferating chondrocyte zone and inhibited chondrocyte hypertrophy and endochondral ossification. Conclusion: The results of this study indicated that, when combined with BMP2, HIF-1α induced MSC differentiation could become a new method of maintaining cartilage phenotypes during cartilage tissue engineering.

scholarly journals Effect of Activated Platelet-Rich Plasma on Chondrogenic Differentiation of Rabbit Bone Marrow-Derived Mesenchymal Stem Cells

2021 ◽  
Author(s):  
Zhen Wang ◽  
Zheng Wang ◽  
Bin Zhang ◽  
Qinghua Zhao ◽  
Yubao Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Chondrogenic Differentiation ◽  
Platelet Rich Plasma ◽  
Cell Counting ◽  
Chondrogenic Medium ◽  
Tissue Repair And Regeneration ◽  
Rabbit Bone

Abstract Background Platelet-rich plasma (PRP) has revealed benefits in tissue repair and regeneration, however, the effect of PRP on proliferation and chondrogenesis of mesenchymal stem cells remains controversial.This study is to evaluate the effect of different concentration PRP on proliferation and chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells(BMSCs). Methods PRP was obtained by centrifugation and activation, in which growth factors and cytokines were detected. BMSCs were isolated from rabbit bone marrow and characterized by flow cytometry. About 5 × 103 BMSCs were cultured in high glucose Dulbecco’s modified Eagle’s medium (HG-DMEM) with 4 different compositions: 10% fetal bovine serum, 5%PRP, 10%PRP, and 15%PRP for consecutive 7 days. Cell counting assays were performed on days 1, 3, 5, and 7 to evaluate the BMSCs proliferation. In chondrogenic differentiation, high-density cell pellets composed of 5 × 105 BMSCs were induced in 4 conditions: commercial chondrogenic medium (control), 5%PRP (HG-DMEM + 5%PRP), 10%PRP (HG-DMEM + 10%PRP), and 15%PRP (HG-DMEM + 15%PRP) for 21 days. The gene expression levels of aggrecan (ACAN), collagen type Ⅱ Alpha 1 (COL2A1), and SRY-Box Transcription Factor 9 (SOX9) in pellets were detected. Histological assessments were performed by morphologically observation and pathological stain. Independent-samples t-test and one-way analysis of variance were used in statistical analyses. Results The concentrations of growth factors and cytokines were elevated in PRP. BMSCs proliferation was enhanced in all groups, and 10% PRP revealed more obvious outcome than the others from day 5. In chondrogenic differentiation, the levels of ACAN, COL2A1, and SOX9 were lower in 3 PRP groups than control, but ACAN and SOX9 was higher in 10% PRP group than 5% and 15%. Histological examinations showed 10% PRP-treated pellets showed more regular appearance, larger size, and abundant extracellular matrix than 5% and 10% groups, but still inferior to commercial chondrogenic medium. Conclusions PRP may enhance the proliferation of rabbit BMSCs, while with limited effect on chondrogenic differentiation compared with commercial chondrogenic medium in pellets culture. Whether optimizing and modifying PRP components would lead to satisfying chondrogenesis of BMSCs, it is deserved furthermore study.

scholarly journals Effect of Activated Platelet-Rich Plasma on Chondrogenic Differentiation of Rabbit Bone Marrow-Derived Mesenchymal Stem Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Zhen Wang ◽  
Zheng Wang ◽  
Bin Zhang ◽  
Qinghua Zhao ◽  
Yubao Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Platelet Rich Plasma ◽  
Cell Counting ◽  
Control Group ◽  
Chondrogenic Medium ◽  
Rabbit Bone ◽  
Fetal Bovine

We aimed to evaluate the effect of activated platelet-rich plasma (PRP) on proliferation and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Six mature male rabbits were included in this study. PRP was obtained by two-step centrifugation from whole blood, and it was activated using CaCl2 solution. BMSCs were isolated and proliferated from bone marrow of rabbits and characterized by flow cytometry. Passage 3 BMSCs were cultured in high-glucose Dulbecco’s modified Eagle’s medium (HG-DMEM) with the four different compositions for consecutive 7 days, including 10% fetal bovine serum, 5% PRP, 10% PRP, and 15% PRP. Cell counting assays were performed to evaluate the cell proliferation of BMSCs. BMSCs ( 5 × 10 5 cells/well in 6-well plates) were induced in four conditions for 21 days to chondrogenic differentiation evaluation, including commercial chondrogenic medium (control), 5% PRP (HG-DMEM+5% PRP), 10% PRP (HG-DMEM+10% PRP), and 15% PRP (HG-DMEM+15% PRP). The gene expression levels of ACAN, COL2A1, and SOX9 in pellets were detected. Morphological and pathological assessments were performed by the blind observer. After purifying, the percentages of cells with CD105(+)/CD34(−) and CD44(+)/CD45(−) were 96.5% and 92.9%, respectively. The proliferation of BMSCs was enhanced in all groups, and 10% PRP revealed more significant outcome than the others from day 5. The levels of ACAN, COL2A1, and SOX9 were lower in the three PRP groups than control group, but the levels of ACAN and SOX9 were higher in 10% PRP group than 5% and 15% PRP groups. Histological examinations showed that 10% PRP-treated pellets had more regular appearance, larger size, and abundant extracellular matrix than 5% or 10% PRP groups, but still inferior to commercial chondrogenic medium. In conclusion, our results show that PRP may enhance the proliferation of rabbit BMSCs. However, PRP have limited effect on chondrogenic differentiation in comparison with commercial chondrogenic medium in pellets culture.

Controlled Differentiation of Mesenchymal Stem Cells into Hyaline Cartilage in miR-140-Activated Collagen Hydrogel

Cartilage ◽  
2021 ◽  
pp. 194760352110476
Author(s):  
Karthikeyan Rajagopal ◽  
Porkizhi Arjunan ◽  
Srujan Marepally ◽  
Vrisha Madhuri
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cartilage Repair ◽  
Chondrogenic Differentiation ◽  
Hyaline Cartilage ◽  
Cartilage Tissue ◽  
Bone Marrow Mscs ◽  
Rt Pcr ◽  
Collagen Hydrogel

Objective Hypertrophic cartilage formation is a major setback in mesenchymal stem cells (MSCs)–mediated cartilage repair, and overcoming it requires optimization of differentiation. Here, we tested the miR-140 activated collagen hydrogel for the chondrogenic differentiation of MSCs and to produce hyaline cartilage. Methods Bone marrow MSCs isolated from 3 patients were pretreated with miR-140 and then chondrogenic differentiated. The 3-dimensional (3D) transfection potential of 5 different transfection reagents (Polyethylenimine, Lipofectamine, TransIT-X2, Amide:Cholesterol-based liposomes [AmC] and AmC pegylated with Tocofersolan [AmCTOC]) was compared and the reagent that showed higher green fluorescent protein (GFP) expression was selected. Finally, the collagen hydrogel was activated using miR-140-transfection complex and sustained delivered to MSCs during chondrogenic differentiation. After differentiation, the outcome was assessed by reverse transcription–polymerase chain reaction (RT-PCR), histology, immunohistochemistry, and compared with scrambled miRNA treated control. Results Pretreatment of MSCs with miR-140 significantly increased the expression of cartilage-specific genes ( COL2A1, SOX9, and ACAN) with reduced hypertrophic chondrocyte ( COL10A1) marker expression and better safranin-O staining than the control. The AmCTOC liposome showed a significant increase in 3D transfection of GFP expressing plasmid than the others. Furthermore, the knockdown of GAPDH using siRNA in HEK cells and expression of GFP mRNA in human bone marrow MSCs confirmed the 3D-transfection efficiency of AmCTOC. The sustained delivery of miR-140 using activated matrix formed a hyaline cartilage-like tissue with minimal COL10A1 expression in RT-PCR and immunohistochemistry. Conclusion Our results demonstrated the therapeutic potential of miR-140-activated hydrogel for MSCs-based cartilage tissue engineering, which could also be used for endogenous stem cells–mediated cartilage repair.

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