Isolation and identification of stem cells in different subtype of cartilage tissue

Xue, Ke; Xia, Wanyao; Zhang, Xiaodie; Qi, Lin; Zhou, Jia; Xu, Peng; Li, Kai

doi:10.1517/14712598.2015.989207

Cited by 20 publications

(19 citation statements)

References 23 publications

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“…CPCs in the auricular cartilage tissue of rabbits (provided by Shanghai Chuansha Breeding Factory, n = 6) were harvested via differential adhesion to fibronectin as described previously [ 31 , 32 ]. Briefly, cells were seeded onto 100-mm plastic petri dishes (pretreated with 10 μg/mL fibronectin overnight at 37 °C) in low-glucose Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA).…”

Section: Methodsmentioning

confidence: 99%

“…After 20 min, the cells were rinsed twice with phosphate-buffered saline (PBS) and cultured in low-glucose DMEM containing 10% fetal bovine serum (FBS, Gibco, USA). BMSCs and chondrocytes from rabbits were isolated and expanded according to previously established methods [ 31 , 33 ]. The isolated cells were cultured in DMEM supplemented with 10% FBS and expanded to passage 2 (P2) to extract exosomes.…”

Section: Methodsmentioning

confidence: 99%

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Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells

et al. 2018

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BackgroundDeveloping cartilage constructed with the appropriate matrix composition and persistent chondrogenesis remains an enduring challenge in cartilage defects. Cartilage progenitor cell (CPC)-based tissue engineering has attracted recent attention because of its strong chondrogenic differentiation capacity. However, due to the lack of a suitable chondrogenic niche, the clinical application of CPC-regenerated cartilage in the subcutaneous environment remains a challenge. In this study, exosomes derived from chondrocytes (CC-Exos) were used to provide the CPC constructs with a cartilage signal in subcutaneous environments for efficient ectopic cartilage regeneration.MethodsRabbit CPC-alginate constructs were prepared and implanted subcutaneously in nude mice. CC-Exos were injected into the constructs at the same dose (30 μg exosomes per 100 μL injection) after surgery and thereafter weekly for a period of 12 weeks. Exosomes derived from bone mesenchymal stem cells (BMSC-Exos) were used as the positive control. The mice in the negative control were administered with the same volume of PBS. At 4 and 12 weeks after implantation, the potential of CC-Exos and BMSC-Exos to promote chondrogenesis and stability of cartilage tissue in a subcutaneous environment were analyzed by histology, immunostaining, and protein analysis. The influences of BMSC-Exos and CC-Exos on chondrogenesis and angiogenic characteristics in vitro were assessed via coculturing with CPCs and human umbilical vein endothelial cells.ResultsThe CC-Exos injection increased collagen deposition and minimized vascular ingrowth in engineered constructs, which efficiently and reproducibly developed into cartilage. The generated cartilage was phenotypically stable with minimal hypertrophy and vessel ingrowth up to 12 weeks, while the cartilage formed with BMSC-Exos was characterized by hypertrophic differentiation accompanied by vascular ingrowth. In vitro experiments indicated that CC-Exos stimulated CPCs proliferation and increased expression of chondrogenesis markers while inhibiting angiogenesis.ConclusionsThese findings suggest that the novel CC-Exos provides the preferable niche in directing stable ectopic chondrogenesis of CPCs. The use of CC-Exos may represent an off-the-shelf and cell-free therapeutic approach for promoting cartilage regeneration in the subcutaneous environment.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells

et al. 2018

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“…BM-derived MSCs express surface markers, including CD90, CD44, CD105, and CD166, and lack hematopoietic lineage markers, CD34 and CD45 (Kala et al, 2012;Xue et al, 2015). A high proportion of cells expressed MSC-specific cell type markers, including CD34, CD44, CD90, and CD105, but did not express CD34, indicating that the cells in this study were of mesenchymal origin and pure.…”

Section: Discussionmentioning

confidence: 69%

Comparative study of neural differentiation of bone marrow mesenchymal stem cells by different induction methods

Zhao

2015

Genet. Mol. Res.

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ABSTRACT. Neurogenic differentiation of bone marrow (BM) mesenchymal stem cells (MSCs) offers a new hope for patients with many neurological disorders. Several chemical inducers are employed to induce BMMSCs differentiation into nerve cells. In the present study, we compared different inducers [2-mercaptoethanol (BME), tretinoin (ATRA), dimethyl sulfoxide/ butylated hydroxyanisole (DMSO/BHA), and indomethacin/3-isobutyl-1-methylxanthine (indomethacin/IBMX)] on the neurogenic differentiation of BMMSCs and aimed to identify a more efficient and safer method. The MSCs were first identified by their ability to adhere to plastic and by the expression of positive (CD44, CD90, and CD105) and negative (CD34) markers assessed by flow cytometry. The efficiency of the neurogenic differentiation was determined by assessing the mRNA and protein expression of nestin, microtubule-associated protein-2 (MAP2), neuron specific enolase (NSE), and glial fibrillary acidic protein (GFAP) by reverse transcription-polymerase chain reaction and western-blot, respectively. The effect of these inducers on cell viability was also evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. This comprehensive study shows that indomethacin/IBMX is better than BME, DMSO/BHA, and ATRA both in terms of efficiency and safety, while BME suppressed the growth and proliferation of MSCs.

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“…Thus, intrinsic regeneration capacity mediated by CPCs may be critical to cartilage degeneration and regeneration [28]. Previously, our and other lab have successfully identified, isolated, and characterized cells with stem or progenitor properties in the articular cartilage of mice models [26,29,30]. CPCs exhibited stem cell characteristics, such as multipotency, clonogenicity, and migratory activity.…”

Section: Discussionmentioning

confidence: 99%

Intra-articular delivery of extracellular vesicles secreted by chondrogenic progenitor cells from MRL/MpJ superhealer mice enhances articular cartilage repair in a mouse injury model

et al. 2020

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Background: Chondrogenic progenitor cells (CPCs) have high self-renewal capacity and chondrogenic potential. Intra-articular delivery of purified mesenchymal stem cells (MSCs) from MRL/MpJ "superhealer" mice increased bone volume during repair and prevents post-traumatic arthritis. Recently, although extracellular vesicles released from MSCs have been used widely for treating OA, the application of extracellular vesicles secreted by CPCs from MRL/ MpJ mice in OA therapy has never been reported. In this study, we evaluated the effects of extracellular vesicles secreted by CPCs from control CBA (CBA-EVs) and MRL/MpJ mice (MRL-EVs) on proliferation and migration of murine chondrocytes. We also determined here if weekly intra-articular injections of CBA-EVs and MRL-EVs would repair and regenerate surgically induced model in mice. Methods: CPC surface markers were detected by flow cytometry. CBA-EVs and MRL-EVs were isolated using an ultrafiltration method. Nanoparticle tracking analysis, transmission electron microscopy, and western blots were used to identify extracellular vesicles. CBA-EVs and MRL-EVs were injected intra-articularly in a mouse model of surgical destabilization of the medial meniscus (DMM)-induced OA, and histological and immunohistochemistry analyses were used to assess the efficacy of exosome injections. We used miRNA-seq analysis to analyze the expression profiles of exosomal miRNAs derived from CBA-EVs as well as MRL-EVs. Cell-counting and scratch assays were used to evaluate the effects of CBA-EVs and MRL-EVs on proliferation and migration of murine chondrocytes, respectively. Meanwhile, a specific RNA inhibitor assesses the roles of the candidate miRNAs in CPC-EV-induced regulation of function of chondrocytes.

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Isolation and identification of stem cells in different subtype of cartilage tissue

Abstract: CSC may become an ideal seeding cell in cartilage tissue engineering, owing to its stemness and chondrogenic characteristics.

Cited by 20 publications

References 23 publications

Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells

Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells

Comparative study of neural differentiation of bone marrow mesenchymal stem cells by different induction methods

Intra-articular delivery of extracellular vesicles secreted by chondrogenic progenitor cells from MRL/MpJ superhealer mice enhances articular cartilage repair in a mouse injury model

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