Magnetic-Based Cell Isolation Technique for the Selection of Stem Cells

Korkusuz, Petek; Köse, Sevil; Yersal, Nilgün; Önen, Selin

doi:10.1007/7651_2018_151

Cited by 8 publications

(6 citation statements)

References 4 publications

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“…After removing red blood cells and dead cells, the isolated single cells were applied to magnetic cell sorting (MACS) according to the method of Korkusuz et al 26 . Pdgfra + cells and Pdgfra − cells were separated using magnetic microbeads conjugated with anti‐Pdgfra (CD140a) antibody (CD140a antibody, anti‐mouse, 130‐101‐905, Miltenyi Biotec, Bergisch Gladbach, Germany).…”

Section: Methodsmentioning

confidence: 99%

Single‐cell RNA sequencing and lipidomics reveal cell and lipid dynamics of fat infiltration in skeletal muscle

You

Chen

et al. 2020

J cachexia sarcopenia muscle

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Background Ageing is accompanied by sarcopenia and intramuscular fat (IMAT) infiltration. In skeletal muscle, fat infiltration is a common feature in several myopathies and is associated with muscular dysfunction and insulin resistance. However, the cellular origin and lipidomic and transcriptomic changes during fat infiltration in skeletal muscle remain unclear. Methods In the current study, we generated a high IMAT‐infiltrated skeletal muscle model by glycerol (GLY) injection. Single‐cell RNA sequencing and lineage tracing were performed on GLY‐injured skeletal muscle at 5 days post‐injection (DPI) to identify the cell origins and dynamics. Lipidomics and RNA sequencing were performed on IMAT‐infiltrated skeletal muscle at 14 DPI (or 17 DPI for the cold treatment) to analyse alterations of lipid compositions and gene expression levels. Results We identified nine distinct major clusters including myeloid‐derived cells (52.13%), fibroblast/fibro/adipogenic progenitors (FAPs) (23.24%), and skeletal muscle stem cells (2.02%) in GLY‐injured skeletal muscle. Clustering and pseudotemporal trajectories revealed six subpopulations in fibroblast/FAPs and 10 subclusters in myeloid‐derived cells. A subpopulation of myeloid‐derived cells expressing adipocyte‐enriched genes and Pdgfra−/Cd68+ cells displayed lipid droplets upon adipogenic induction, indicating their adipogenic potential. Lipidomic analysis revealed the changes of overall lipid classes composition (e.g. triglycerides (TAGs) increased by 19.3 times, P = 0.0098; sulfoquinovosyl diacylglycerol decreased by 83%, P = 0.0056) and in the distribution of lipids [e.g. TAGs (18:2/18:2/22:6) increased by 181.6 times, P = 0.021] between GLY‐group and saline control. RNA‐seq revealed 1847 up‐regulated genes and 321 down‐regulated genes and significant changes in lipid metabolism‐related pathways (e.g. glycerolipid pathway and glycerophospholipid pathway) in our model of GLY‐injured skeletal muscle. Notably, short‐term cold exposure altered fatty acid composition (e.g. saturated fatty acid decreased by 6.4%, P = 0.058) in fat‐infiltrated muscles through directly affecting lipid metabolism pathways including PI3K–AKT and MAPK signalling pathway. Conclusions Our results showed that a subpopulation of myeloid‐derived cells may contribute to IMAT infiltration. GLY‐induced IMAT infiltration changed the lipid composition and gene expression profiles. Short‐term cold exposure might regulate lipid metabolism and its related signalling pathways in fat‐infiltrated muscle. Our study provides a comprehensive resource describing the molecular signature of fat infiltration in skeletal muscle.

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Section: Methodsmentioning

confidence: 99%

Single‐cell RNA sequencing and lipidomics reveal cell and lipid dynamics of fat infiltration in skeletal muscle

You

Chen

et al. 2020

J cachexia sarcopenia muscle

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“…Magnetic beads are conjugated to highly specific monoclonal antibodies that recognize CCSC marker on the surface of cells of interest. Then, the heterogeneous suspension of cells is passed through a separation column, in a magnetic field, to retain the cells labeled with magnetic beads and antibodies [ 45 ]. By switching off the magnetic field, target cells will be eluted.…”

Section: Colorectal Cancer Stem Cellsmentioning

confidence: 99%

“…MACS is a fast and easy method of cell separation, especially for the isolation of CCSCs that represent a small cell population in the tumor mass. However, MACS is only a mono-parameter separation method that requires cell labelling and is unable to separate cells based on the variable expression of markers [ 44 , 45 ].…”

Section: Colorectal Cancer Stem Cellsmentioning

confidence: 99%

The Role of Cancer Stem Cells in Colorectal Cancer: From the Basics to Novel Clinical Trials

Hervieu

Christou

Battu

et al. 2021

Cancers

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The treatment options available for colorectal cancer (CRC) have increased over the years and have significantly improved the overall survival of CRC patients. However, the response rate for CRC patients with metastatic disease remains low and decreases with subsequent lines of therapy. The clinical management of patients with metastatic CRC (mCRC) presents a unique challenge in balancing the benefits and harms while considering disease progression, treatment-related toxicities, drug resistance and the patient’s overall quality of life. Despite the initial success of therapy, the development of drug resistance can lead to therapy failure and relapse in cancer patients, which can be attributed to the cancer stem cells (CSCs). Thus, colorectal CSCs (CCSCs) contribute to therapy resistance but also to tumor initiation and metastasis development, making them attractive potential targets for the treatment of CRC. This review presents the available CCSC isolation methods, the clinical relevance of these CCSCs, the mechanisms of drug resistance associated with CCSCs and the ongoing clinical trials targeting these CCSCs. Novel therapeutic strategies are needed to effectively eradicate both tumor growth and metastasis, while taking into account the tumor microenvironment (TME) which plays a key role in tumor cell plasticity.

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“…The more calibrated samples in the runner of the disc, the large number of isolated samples will be obtained. This helps to improve the accuracy of detection [15]. Therefore, the pattern of the disc was designed as shown in Figure 1a.…”

Section: Design the Pattern Of The Discmentioning

confidence: 99%

How to Control the Microfluidic Flow and Separate the Magnetic and Non-Magnetic Particles in the Runner of a Disc

2021

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Biochips play an important role in both medical and food industry safety testing. Moreover, magnetic activated cell sorting is a well-established technology for biochip development. However, biochips need to be manufactured by precision instruments, resulting in the high cost of biochips. Therefore, this study used magnetic-activation and mechanics theories to create a novel disc that could manipulate the microfluidic flow, mixing, reaction, and separation on the runner of the disc. The goal of the research was to apply in the field of biomedical detection systems to reduce the cost of biochips and simplify the operation process. The simulation and experimental investigation showed that the pattern of the reaction chamber was stomach-shaped and the reservoir chamber was rectangular-shaped on the disc. The microfluid could be controlled to flow to the reaction chamber from the buffer and sample chamber when the disc spun at 175~200 rpm within three minutes. This was defined as the first setting mode. The microfluid could then be controlled to flow to the reservoir chamber from the reaction chamber when the disc spun at 225 rpm within five to ten minutes. This was defined as the second setting mode. This verified that the pattern design of the disc was optimized for control of the microfluid flow, mixing, reaction, and separation in the runner of the disc by different setting modes.

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Magnetic-Based Cell Isolation Technique for the Selection of Stem Cells

Cited by 8 publications

References 4 publications

Single‐cell RNA sequencing and lipidomics reveal cell and lipid dynamics of fat infiltration in skeletal muscle

Single‐cell RNA sequencing and lipidomics reveal cell and lipid dynamics of fat infiltration in skeletal muscle

The Role of Cancer Stem Cells in Colorectal Cancer: From the Basics to Novel Clinical Trials

How to Control the Microfluidic Flow and Separate the Magnetic and Non-Magnetic Particles in the Runner of a Disc

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