Methods of Cell Purification: A Critical Juncture for Laboratory Research and Translational Science

Amos, Peter J.; Çağavı, Esra; Qyang, Yibing

doi:10.1159/000331390

Cited by 29 publications

(26 citation statements)

References 196 publications

(134 reference statements)

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“…The cytokine pattern could evidently be different from the ones produced by the blood cells. However, the use of these cells often requires invasive techniques to obtain them and their purification, for subsequent activation, is more labour-intensive than blood cells [27]. It is also useful to stress that the presence of a SNP in TRAF3IP2 rs33980500, as shown in this case report, should be present in all cell types using this signalling pathway [28,29].…”

Section: Discussionmentioning

confidence: 89%

The use of leukocytes’ secretome to individually target biological therapy in autoimmune arthritis: a case report

Poubelle

Pagé

Longchamps

et al. 2019

Clinical & Translational Med

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Background Biological agents have allowed remarkable improvement in controlling autoimmune arthropathies, although none of the numerous biologics readily available represent a universal treatment standard. Moreover, classical and genetic predictors are currently unsatisfactory to predict individual response to a biologic, and the best treatment selection is still based on a trial-and-error approach. Here, we report a clinical case demonstrating the usefulness of examining the leukocytes’ secretome of patients. We set up and standardized a protocol that examines a patient’s immune responses to establish the secretome of the blood mononuclear leukocytes and personalize the biotherapy. Case presentation A 24-year-old woman was diagnosed with active early rheumatoid arthritis. The initial treatment regimen (prednisone, methotrexate, hydroxychloroquine, naproxen) was inefficient, as well as the anti-TNF adalimumab. The diagnosis was revised as possible rheumatoid arthritis-like psoriatic arthritis and adalimumab was replaced by abatacept (IgG1 Fc-CTLA-4) to no avail. Five years later, abatacept was replaced by the anti-IL-12/IL-23 ustekinumab with no objective control over the symptoms. The patient was thus enrolled in a prospective study based on the quantification of cytokines secreted by peripheral blood leukocytes stimulated with well-known immune activators of pattern recognition receptors and cytokine signalling. The results of this study revealed that plasma concentrations of cytokines were similar between the patient and healthy donors. In comparison to leukocytes from healthy donors, the patient’s secretome showed a unique overproduction of IL-6. The anti-IL-6 receptor tocilizumab was, therefore, administered with a rapid improvement of her active psoriatic arthritis that remained dependent on low prednisone dosage. Clinical parameters progressively returned to normal levels and her quality of life was greatly improved, despite the major delay to begin the present personalized treatment. Conclusions An efficient way to effectively treat patients with complex autoimmune arthropathies, and avoid irreversible disability, is to know their leukocytes’ secretome to identify abnormally secreted cytokines and personalize their biotherapy, as exemplified by this case report.

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Section: Discussionmentioning

confidence: 89%

The use of leukocytes’ secretome to individually target biological therapy in autoimmune arthritis: a case report

Poubelle

Pagé

Longchamps

et al. 2019

Clinical & Translational Med

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“…Magnetic micro/nanoparticle-based cell sorting is a second common approach which utilizes antibody specificity to separate cells of interest from bulk cell mixtures and is often inexpensive and easy to use. 1,6,7 However, magnetic interaction between microparticles and some sensitive cell populations such as stem and progenitor cells is known to have negative impact on cell viability, phenotypic identity, and cell function. [8][9][10][11] Further, the high concentrations of small particles (<2 microns) commonly used in magnetic isolation risk significant cell stress due to particle internalization.…”

mentioning

confidence: 99%

Cell Isolation and Recovery Using Hollow Glass Microspheres Coated with Nanolayered Films for Applications in Resource-Limited Settings

Dong

Ahrens

et al. 2017

ACS Appl. Mater. Interfaces

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Established cell isolation and purification techniques such as fluorescence-activated cell sorting (FACS), isolation through magnetic micro/nanoparticles, and recovery via microfluidic devices have limited application as disposable technologies appropriate for point-of-care use in remote areas where lab equipment as well as electrical, magnetic, and optical sources are restricted. We report a simple yet effective method for cell isolation and recovery that requires neither specialized lab equipment nor any form of power source. Specifically, self-floating hollow glass microspheres were coated with an enzymatically degradable nanolayered film and conjugated with antibodies to allow both fast capture and release of subpopulations of cells from a cell mixture. Targeted cells were captured by the microspheres and allowed to float to the top of the hosting liquid, thereby isolating targeted cells. To minimize nonspecific adhesion of untargeted cells and to enhance the purity of the isolated cell population, an antifouling polymer brush layer was grafted onto the nanolayered film. Using the EpCAM-expressing cancer cell line PC-3 in blood as a model system, we have demonstrated the isolation and recovery of cancer cells without compromising cell viability or proliferative potential. The whole process takes less than 1 h. To support the rational extension of this platform technology, we introduce extensive characterization of the critical design parameters: film formation and degradation, grafting with a poly(ethylene glycol) (PEG) sheath, and introducing functional antibodies. Our approach is expected to overcome practical hurdles and provide viable targeted cells for downstream analyses in resource-limited settings.

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“…The purification of different types of cultured cells is critical in various biomedical fields, including basic research, drug development, and cell therapy. Conventionally, fluorescenceactivated cell sorting (FACS), affinity beads (e.g., magnetic-activated cell sorting (MACS)), gradient centrifugation, and elutriation have been used for cell purification 1 . However, these technologies are essentially aimed at floating cells in suspension.…”

Section: Introductionmentioning

confidence: 99%

Automated Adherent Cell Elimination by a High-Speed Laser Mediated by a Light-Responsive Polymer

Hayashi

Matsumoto²,

Kumagai

et al. 2018

Preprint

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Conventional cell handling and sorting methods require manual dissociation, which decreases cell quality and quantity and is not well suited for monitoring applications. To purify adherent cultured cells, in situ cell purification technologies that are high throughput and can be utilized in an on-demand manner are expected. Previous demonstrations using direct laser-mediated cell elimination revealed only limited success in terms of their usability and throughput. Here, we developed a Laser-Induced, Light-responsive-polymer-Activated, Cell Killing (LILACK) system that enables high-speed and on-demand adherent cell sectioning and purification. This system employs a visible laser beam, which does not kill cells directly, but induces local heat production through the trans-cis-trans photo-isomerization of azobenzene moieties in only the irradiated area of a light-responsive thin layer. Using this system in each passage for sectioning, human induced pluripotent stem cells (hiPSCs) were maintained their pluripotency and self-renewal during long-term culture. Furthermore, combined with deep machine-learning analysis on fluorescent and phase contrast images, a label-free and automatic cell processing system was developed by eliminating unwanted spontaneously differentiated cells in undifferentiated hiPSC culture conditions.

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Methods of Cell Purification: A Critical Juncture for Laboratory Research and Translational Science

Cited by 29 publications

References 196 publications

The use of leukocytes’ secretome to individually target biological therapy in autoimmune arthritis: a case report

The use of leukocytes’ secretome to individually target biological therapy in autoimmune arthritis: a case report

Cell Isolation and Recovery Using Hollow Glass Microspheres Coated with Nanolayered Films for Applications in Resource-Limited Settings

Automated Adherent Cell Elimination by a High-Speed Laser Mediated by a Light-Responsive Polymer

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