A fully automated primary neuron purification system using continuous centrifugal microfluidics

Intisar, Aseer; Lee, Seung Joon; Kim, Yu-Gyeong; Kim, Woon‐Hae; Kim, Min Young; Kim, Jong Man; Lee, Jungmin; Mo, Yun Jeoung; Kim, Yu Seon; Kim, Seung‐Hoon; Lee, Yun‐Il; Kim, Minseok S.

doi:10.1039/d2lc00381c

Cited by 4 publications

(6 citation statements)

References 34 publications

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“…Since they must be stopped in order to open a ferrowax valve, the separated fluidic layers via centrifugation are mixed, failing the extraction of a specific fluidic layer . To overcome this challenge, the CCM technique was implemented. , This method synchronizes the rotation of the laser and disc during the valve opening step (Figure B). Therefore, the disc rotation does not need to be stopped completely, enabling the ultra-thin buffy coat to be maintained throughout the whole process.…”

Section: Resultsmentioning

confidence: 99%

“…36 To overcome this challenge, the CCM technique was implemented. 35,38 This method synchronizes the rotation of the laser and disc during the valve opening step (Figure 2B). Therefore, the disc rotation does not need to be stopped completely, enabling the ultra-thin buffy coat to be maintained throughout the whole process.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…In this study, we introduce a fully automated continuous centrifugal microfluidics-based NK disc (CCM-NKD) for NK cell isolation. The centrifugal microfluidic system has been investigated and applied in various fields, including the isolation of rare cells such as CTCs , and trophoblasts, as well as primary neuron cell isolation and nucleic acid testing . The CCM-NKD implements a negative selection strategy, eliminating non-NK cells to increase the NK cell concentration.…”

Section: Introductionmentioning

confidence: 99%

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Fully Automated Continuous Centrifugal Microfluidics Isolates Natural Killer Cells with High Performance and Minimal Stress

Yang

Kim²,

Intisar

et al. 2023

Anal. Chem.

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Natural killer (NK) cells are a part of the innate immune system, providing the first line of defense against cancer cells and pathogens at an early stage. Hence, they are attracting attention as a valuable resource for allogeneic cell immunotherapy. However, NK cells exist with limited proportion in blood, and obtaining sufficient clinical-grade NK cells with highly viable and minimal stress is critical for successful immune cell therapy. Conventional purification methods via immunoaffinity or density gradient centrifugation had several limitations in yield, purity, and cellular stress, which might cause an increased risk for graft versus host disease and reduced efficacy due to NK cell malfunction, exhaustion, and apoptosis. Moreover, reducing the variations of isolation performance caused by the manual process is another unmet need for uniform quality of the living drug. Here, an automated system using an NK disc (NKD) based on continuous centrifugal microfluidics (CCM) technology was developed to isolate NK cells from whole blood with high yield, purity, reproducibility, and low stress. The CCM technology, which operates fluidic manipulation under disc rotation, enabled precise extraction of the ultra-thin target fluid layer generated by blood centrifugation. Compared to the conventional manual method, the CCM-NKD isolated NK cells with higher yield (recovery rate) and purity, while maintaining better reproducibility. Furthermore, since the CCM-NKD maintained substantially milder centrifugation conditions (120 ×g for 10 min) compared to the conventional approach (1200 ×g for 20 min), it showed reduced cellular stress and increased antioxidant capacity in the isolated NK cells. Based on the results, the CCM-NKD is expected to be a useful tool to provide highly intact and viable cell weapons for successful immune cell therapy.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Fully Automated Continuous Centrifugal Microfluidics Isolates Natural Killer Cells with High Performance and Minimal Stress

Yang

Kim²,

Intisar

et al. 2023

Anal. Chem.

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“…For example, Intisar et al. [100] used a fully automated lab‐on‐a‐disc device to isolate neuronal cells by the combination of density gradient centrifugation and immunomagnetic separation. This on‐disc multifield assisted extraction speeds up the 1‐wk bench‐top preparation process to only 13 min, which corresponds to an 800‐fold faster process.…”

Section: Microfluidic Sample Preparation Techniquesmentioning

confidence: 99%

“…The on-disc sample handling is implemented by a centrifugal force and the various valves are involved in sample/solution transfer between different sample preparation processes [95,96]. These disc devices are most frequently used for protein [38,97], genes [65,98], bacteria [64,99], and cell [100,101] analysis containing multiple processes. Some of these discs are employed as user-friendly sample preparation equipment for the off-line detection, such as LC-MS/MS [38,97].…”

Section: Lab-on-a-chip Sample Preparation Techniquesmentioning

confidence: 99%

Recent progress of microfluidic sample preparation techniques

Xia

2023

J of Separation Science

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Sample preparation turns out to be one of the important procedures in complex sample analysis by affecting the accuracy, selectivity, and sensitivity of analytical results. However, the majority of the conventional sample preparation techniques still suffer from time‐consuming and labor‐intensive operations. These shortcomings can be addressed by reforming the sample preparation process in a microfluidic manner. Inheriting the advantages of rapid, high efficiency, low consumption, and easy integration, microfluidic sample preparation techniques receive increasing attention, including microfluidic phases separation, microfluidic field‐assisted extraction, microfluidic membrane separation, and microfluidic chemical conversion. This review overviews the progress of microfluidic sample preparation techniques in the last 3 years based on more than 100 references, we highlight the implementation of typical sample preparation methods in the formats of microfluidics. Furthermore, the challenges and outlooks of the application of microfluidic sample preparation techniques are discussed.

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One-step on-chip preparation of nanoparticle-conjugated red blood cell carriers

Xu,

You,

Zhang

et al. 2025

Colloids and Surfaces B: Biointerfaces

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A fully automated primary neuron purification system using continuous centrifugal microfluidics

Abstract: Non-neuronal cells are depleted using immunomagnetic separation and density-gradient centrifugation, resulting in purified primary neurons, obtained 800× faster, while maintaining superior viability and purity, compared to the conventional method.

Cited by 4 publications

References 34 publications

Fully Automated Continuous Centrifugal Microfluidics Isolates Natural Killer Cells with High Performance and Minimal Stress

Fully Automated Continuous Centrifugal Microfluidics Isolates Natural Killer Cells with High Performance and Minimal Stress

Recent progress of microfluidic sample preparation techniques

One-step on-chip preparation of nanoparticle-conjugated red blood cell carriers

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