Magnetic-Activated Cell Sorting Using Coiled-Coil Peptides: An Alternative Strategy for Isolating Cells with High Efficiency and Specificity

Shen, Ming; Olsthoorn, René C. L.; Zeng, Ye; Bakkum, Thomas; Kros, Alexander; Boyle, Aimee L.

doi:10.1021/acsami.0c22185

Cited by 17 publications

(10 citation statements)

References 38 publications

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“…Impaired proliferation of the stem cells (observation) led to a rapid decrease in DSC frequency and takeover of the remaining fibroblasts. It is known that magnetic particles can lead to adverse effects such as influencing cell phenotype, function [ 36 , 37 ], and viability [ 38 , 39 , 40 ]. This indicates that the DSCs might have been affected by the antibodies and RapidSpheres™, which they were still attached to.…”

Section: Resultsmentioning

confidence: 99%

Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts

Starzonek

Mhamdi-Ghodbani

Henning

et al. 2023

Cells

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Dermal stem cells (DSCs), which are progenitor cells of melanocytes, are isolated from human foreskin and cultivated as mixed cultures containing both DSCs and fibroblasts in varying proportions. These contaminating fibroblasts may have an impact on the results of experimental studies and are a serious limitation for certain applications. The aim of the present study was to purify or enrich DSCs—an indispensable step towards future investigations. Applying different methods, we demonstrated that highly enriched DSCs with a good recovery rate can be obtained through positive selection with MACS® immunomagnetic cell sorting. These DSCs remain vital and proliferate constantly in culture, maintaining a high level of purity after enrichment. Other approaches such as treatment with Geneticin or selective detachment were not suitable to purify DSC-fibroblast co-cultures. Overall, enriched DSCs represent a novel and unique model to study the effects of UV radiation on the differentiation of DSCs into melanocytes and their potential relevance in the genesis of malignant melanoma.

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Section: Resultsmentioning

confidence: 99%

Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts

Starzonek

Mhamdi-Ghodbani

Henning

et al. 2023

Cells

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“…Several cell purification methods have been established to collect cells with specific characteristics. Typical methods are (i) magnetic-activated cell sorting (MACS) [ 44 , 45 , 46 ], (ii) fluorescence-activated cell sorting (FACS) [ 47 , 48 ], and (iii) cell selection by genetic editing with antibiotic resistance genes and antibiotic treatment (e.g., puromycin, neomycin, G418, and zeocin) [ 49 , 50 , 51 , 52 , 53 ]. In both the MACS and FACS methods, antibody binding on the targeting cells is essential, which indicates the possibility of antibody contamination on the final product of the specific cells, and antibody contamination in the final product is not desirable for clinical usage of the specific cells.…”

Section: Discussionmentioning

confidence: 99%

Purification of Colon Carcinoma Cells from Primary Colon Tumor Using a Filtration Method via Porous Polymeric Filters

et al. 2021

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Cancer stem cells (CSCs) or cancer-initiating cells (CICs) are key factors for tumor generation and metastasis. We investigated a filtration method to enhance CSCs (CICs) from colon carcinoma HT-29 cells and primary colon carcinoma cells derived from patient colon tumors using poly(lactide-co-glycolic acid)/silk screen (PLGA/SK) filters. The colon carcinoma cell solutions were permeated via porous filters to obtain a permeation solution. Then, the cell cultivation media were permeated via the filters to obtain the recovered solution, where the colon carcinoma cells that adhered to the filters were washed off into the recovered solution. Subsequently, the filters were incubated in the culture media to obtain the migrated cells via the filters. Colon carcinoma HT-29 cells with high tumorigenicity, which might be CSCs (CICs), were enhanced in the cells in the recovered solution and in the migrated cells based on the CSC (CIC) marker expression, colony-forming unit assay, and carcinoembryonic antigen (CEA) production. Although primary colon carcinoma cells isolated from colon tumor tissues contained fibroblast-like cells, the primary colon carcinoma cells were purified from fibroblast-like cells by filtration through PLGA/SK filters, indicating that the filtration method is effective in purifying primary colon carcinoma cells.

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“…The lipopeptide-based fusion system CPE/CPK has already been used in many applications, such as cell sorting, liposome fusion, ,, liposome cell fusion, ,, lipid nanoparticle cell fusion, cell–cell fusion, and anion transporter delivery to membranes …”

Section: Fusogenic Peptidesmentioning

confidence: 99%

“…Besides receptor targeting, peptides can mediate the cellular uptake of functionalized lipid-based nanoparticles via a biomimetic mechanism using a coiled-coil interaction. Examples showed that they can be easily inserted into cells, liposomes, vesicles, and lipid nanoparticles (LNPs). ,, By modifying cells and vehicles, it could be shown that the uptake was drastically increased and selectivity over modified and nonmodified cells was achieved.…”

Section: Lipid-based Nanoparticles Functionalized With Fusogenic Pept...mentioning

confidence: 99%

Lipid-Based Nanoparticle Functionalization with Coiled-Coil Peptides for In Vitro and In Vivo Drug Delivery

Aschmann,

Knol,

Kros

2024

Acc. Chem. Res.

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Metrics & MoreArticle Recommendations CONSPECTUS: For the delivery of drugs, different nanosized drug carriers (e.g., liposomes, lipid nanoparticles, and micelles) have been developed in order to treat diseases that afflict society. Frequently, these vehicles are formed by the self-assembly of small molecules to encapsulate the therapeutic cargo of interest. Over decades, nanoparticles have been optimized to make them more efficient and specific to fulfill tailor-made tasks, such as specific cell targeting or enhanced cellular uptake. In recent years, lipid-based nanoparticles in particular have taken center stage; however, off-targeting side effects and poor endosomal escape remain major challenges since therapies require high efficacy and acceptable toxicity.To overcome these issues, many different approaches have been explored to make drug delivery more specific, resulting in reduced side effects, to achieve an optimal therapeutic effect and a lower required dose. The fate of nanoparticles is largely dependent on size, shape, and surface charge. A common approach to designing drug carriers with targeting capability is surface modification. Different approaches to functionalize nanoparticles have been investigated since the attachment of targeting moieties plays a significant role in whether they can later interact with surface-exposed receptors of cells. To this end, various strategies have been used involving different classes of biomolecules, such as small molecules, nucleic acids, antibodies, aptamers, and peptides. Peptides in particular are often used since there are many receptors overexpressed in different specific cell types. Furthermore, peptides can be produced and modified at a low cost, enabling high therapeutic screening. Cell-penetrating peptides (CPPs) and cell-targeting peptides (CTPs) are frequently used for this purpose. Less studied in this context are fusogenic coiled-coil peptides.Lipid-based nanoparticles functionalized with these peptides are able to avoid the endolysosomal pathway; instead such particles can be taken up by membrane fusion, resulting in increased delivery of payload. Furthermore, they can be used for targeting cells/organs but are not directed at surface-exposed receptors. Instead, they recognize complementary peptide sequences, facilitating their uptake into cells.In this Account, we will discuss peptides as moieties for enhanced cytosolic delivery, targeted uptake, and how they can be attached to lipid-based nanoparticles to alter their properties. We will discuss the properties imparted to the particles by peptides, surface modification approaches, and recent examples showing the power of peptides for in vitro and in vivo drug delivery. The main focus will be on the functionalization of lipid-based nanoparticles by fusogenic coiled-coil peptides, highlighting the relevance of this concept for the development of future therapeutics.

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Magnetic-Activated Cell Sorting Using Coiled-Coil Peptides: An Alternative Strategy for Isolating Cells with High Efficiency and Specificity

Cited by 17 publications

References 38 publications

Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts

Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts

Purification of Colon Carcinoma Cells from Primary Colon Tumor Using a Filtration Method via Porous Polymeric Filters

Lipid-Based Nanoparticle Functionalization with Coiled-Coil Peptides for In Vitro and In Vivo Drug Delivery

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