• 제목/요약/키워드: Cell Sorting

검색결과 136건 처리시간 0.031초

세포 크기 차이를 이용한 유세포 분석을 통한 인간배아줄기세포 유래 기능성 혈관세포의 확립 (Establishment of Functional Cells for Vascular Defect Disease from Human Embryonic Stem Cell via Region Sorting Depending on Cell Volume)

  • 이지혜;김주미;정형민;채정일
    • 한국미생물·생명공학회지
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    • 제39권4호
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    • pp.364-373
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    • 2011
  • 인간배아줄기세포는 인간배아줄기세포가 가지는 전 분화능 등의 특이적 특성으로 인해 재생의학 분야에서 세포 치료제의 근원으로 널리 각광받고 있다. 그러나, 미분화 상태의 인간배아줄기세포를 세포치료제로 이용하기 위해서는 인간배아줄기세포 주 유래 기능성 세포를 확립이 반드시 요구된다. 본 연구에서는, 미분화 상태의 인간배아줄기세포주로부터 기능성 세포의 확립을 위해, 혈관계통의 세포로 분화를 유도하였으며, 분화 유도 후 세포의 크기 차이를 이용하여 특정 세포군 만을 분리하여 그 기능성을 비교 분석하였다. 그 결과, VEGF를 이용하여 분화 시킨 세포군에서 약 10%의 PECAM 양성 세포군을 확인할 수 있었으며, 분리 및 세포 이식을 위해 세포를 단일 세포군으로 만들었다. 단일 세포군의 형성 후, 유세포 분석기를 이용한 세포 분리 기법을 이용하여 FCS를 기준으로 한 세포 크기의 차이를 이용하여 특정 세포군 만을 분리하여, 하지 허혈 동물 모델로의 이식을 통해, 비 분리 세포군과 치료 효능을 비교 분석을 실시하였다. 세포 이식 4주 후, 혈류량 복구율이 FSC 기준 분리 군의 경우 54%, 비 분리군의 경우 17%를 보이는 것을 확인하였다. 이 결과는, 초기 분화 유도 후 세포 크기차이를 이용한 세포 분리법이 기능성 세포 획득에 이용될 수 있음을 시사한다. 이와 같은 방법을 통해 다양한 종류의 기능성 세포 분리에 이용될 수 있을 것이라 생각된다.

형광 리포터를 활용한 효모 단백질 잡종 기법 개발 (Yeast two-hybrid assay with fluorescence reporter)

  • 박성균;서수련;황병준
    • 미생물학회지
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    • 제55권3호
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    • pp.199-205
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    • 2019
  • Yeast two-hybrid는 특정 단백질에 대한 상호작용 파트너 단백질의 선별을 위한 방법으로 개발되었다. 하지만 대규모 단백질 상호작용체 분석을 수행하기에 요구되는 노동과 대량의 한천배지 사용에 따른 문제에 의해 널리 사용되지 못하고 있다. 따라서 본 연구에서는 새로운 리포터 시스템을 yeast two-hybrid 방법에 도입하여 fluorescence-activated cell sorting (FACS) 또는 magnetic-activated cell sorting (MACS)를 이용하여 상호작용 파트너 단백질을 포함하는 효모 클론을 손쉽게 선별할 수 있도록 하였다. 새로운 리포터 시스템은 c-myc 항원 결정기가 총 10번 반복되는 형태로 효모 표면에 발현되도록 하였으며, p53과 SV40 T항원을 이용한 실험을 통하여 리포터 단백질의 정상적인 발현을 flow cytometry 분석을 통하여 확인하였다. 따라서, 새로운 리포터 시스템을 도입한 yeast two-hybrid 방법은 대규모 상호작용체 분석을 위해 필요한 노력을 현저히 줄일 수 있을 것으로 기대한다.

Microfluidic Devices for Cell Analysis

  • Bachman, Mark;Li, G.P.
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2009년도 추계학술발표대회
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    • pp.3.2-3.2
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    • 2009
  • Microfluidics and BioMEMStechnology has increasingly been used as a tool for studying small volumes oftissue and even individual cells. One of the most important benefits ofmicrofluidic technology is the potential to build devices that analyze and sortmammalian cells. The "sorting problem" typically requires that a fewcells be selected and isolated from a larger population of hundreds, thousandsor even millions of other cells. For example, cancer tumor cells may resideamong a large population of healthy cells, but it would be of great interest toidentify, isolate and study only the cancer cells. In another application, onemay want to determine the number of white blood cells within a sample of blood.We have developed microfluidic devices that enable researchers to select cellsfrom a population by a variety of methods, including antibody staining,dielectrophoretic selection, and physical size selection. These devices haveapplications in cancer research where cancer cells must be identified fromnormal tissue, but where only small samples of tissue are available. In thistalk, we will present some of our microfluidic cell sorting devices, discusstheir physical principles, and their use in biological applications.

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ATM 교환을 위한 비용 효율적인 동적 결함내성 bitonic sorting network (A Cost-Effective Dynamic Redundant Bitonic Sorting Network for ATM Switching)

  • 이재동;김재홍;최홍인
    • 한국정보처리학회논문지
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    • 제7권4호
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    • pp.1073-1081
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    • 2000
  • This paper proposes a new fault-tolerant technique for bitonic sorting networks which can be used for designing ATM switches based on Batcher-Banyan network. The main goal in this paper is to design a cost-effective fault-tolerant bitonic sorting network. In order to recover a fault, additional comparison elements and additional links are used. A Dynamic Redundant Bitonic Sorting (DRBS) network is based on the Dynamic Redundant network and can be constructed with several different variations. The proposed fault-tolerant sorting network offers high fault-tolerance; low time delays; maintenance of cell sequence; simple routing; and regularity and modularity.

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Long-term exposure to gefitinib differentially regulates the endosomal sorting complex required for transport machinery, which accelerates the metastatic potential of oral squamous cell carcinoma cells

  • Mi Seong Kim;Min Seuk Kim
    • International Journal of Oral Biology
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    • 제48권1호
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    • pp.1-7
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    • 2023
  • Oral squamous cell carcinoma (OSCC), which accounts for approximately 90% of oral cancers, has a high rate of local recurrence and a poor prognosis despite improvements in treatment. Exosomes released from OSCC cells promote cell proliferation and metastasis. Although it is clear that the biogenesis of exosomes is mediated by the endosomal sorting complex required for transport (ESCRT) machinery, the gene expression pattern of ESCRT, depending on the cell type, remains elusive. The exosomal release from the human OSCC cell lines, HSC-3 and HSC-4, and their corresponding gefitinib-resistant sub-cell lines, HSC-3/GR and HSC-4/GR, was assessed by western blot and flow cytometry. The levels of ESCRT machinery proteins, including Hrs, Tsg101, and Alix, and whole-cell ubiquitination were evaluated by western blot. We observed that the basal level of exosomal release was higher in HSC-3/GR and HSC-4/GR cells than in HSC-3 and HSC-4 cells, respectively. Long-term gefitinib exposure of each cell line and its corresponding gefitinib-resistant sub-cell line differentially induced the expression of the ESCRT machinery. Furthermore, whole-cell ubiquitination and autophagic flux were shown to be increased in gefitinib-treated HSC-3 and HSC-4 cells. Our data indicate that the expression patterns of the ESCRT machinery genes are differentially regulated by the characteristics of cells, such as intracellular energy metabolism. Therefore, the expression patterns of the ESCRT machinery should be considered as a key factor to improve the treatment strategy for OSCC.

In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju;Lee Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • 제16권7호
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    • pp.1149-1153
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    • 2006
  • In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.

Inertial Microfluidics-Based Cell Sorting

  • Kim, Ga-Yeong;Han, Jong-In;Park, Je-Kyun
    • BioChip Journal
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    • 제12권4호
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    • pp.257-267
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    • 2018
  • Inertial microfluidics has attracted significant attention in recent years due to its superior benefits of high throughput, precise control, simplicity, and low cost. Many inertial microfluidic applications have been demonstrated for physiological sample processing, clinical diagnostics, and environmental monitoring and cleanup. In this review, we discuss the fundamental mechanisms and principles of inertial migration and Dean flow, which are the basis of inertial microfluidics, and provide basic scaling laws for designing the inertial microfluidic devices. This will allow end-users with diverse backgrounds to more easily take advantage of the inertial microfluidic technologies in a wide range of applications. A variety of recent applications are also classified according to the structure of the microchannel: straight channels and curved channels. Finally, several future perspectives of employing fluid inertia in microfluidic-based cell sorting are discussed. Inertial microfluidics is still expected to be promising in the near future with more novel designs using various shapes of cross section, sheath flows with different viscosities, or technologies that target micron and submicron bioparticles.

Analysis of Membrane Integrity, DNA Fragmentation and Mitochondrial Function in Pig Spermatozoa Sorted by Flowcytometer

  • Kim, In-Cheul;Han, Deug-Woo;Lee, Sung-Won;Ryu, Jae-Weon;Choi, Eun-Ji;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • 제32권2호
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    • pp.123-126
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    • 2008
  • The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

Development and Performance Evaluation of Falling-type Dried-Persimmon Weight Sorting System Utilizing Load Cell

  • Lim, Jongguk;Kim, Giyoung;Mo, Changyeun;Choi, Inchul
    • Journal of Biosystems Engineering
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    • 제40권4호
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    • pp.327-334
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    • 2015
  • Purpose: A falling-type weight sorter equipped with a load cell was developed to sort lightweight dried persimmons. The performance of the sorter was also evaluated. Methods: The electronic weight sorter for dried persimmon comprises a feeder part, a weight-measurement part, an indicator part, a carrier cup, a discharging part, and a driving part. The weight setting and zero-point adjustment are performed digitally for the convenience of users. For the experimental trials, 228 rubber-clay specimens (representative of dried persimmons) in the weight range of 24.73~99.56 g were manufactured for use in experiments to evaluate the performance of the sorter. Results: The average error of the weight measurements from three experimental trials was 1.655%, with a bias of -0.492 g, a root-mean-square error (RMSE) of ${\pm}0.808g$, and a coefficient of determination ($R^2$ ) of 0.997. Conclusions: The load-cell-based electronic dried-persimmon weight sorter developed in this study facilitates effective, precise, and convenient sorting of dried persimmons.