• Title/Summary/Keyword: Whole blood cells

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Microsystems for Whole Blood Purification and Electrophysiological Analysis

  • Han, Arum;Han, Ki-Ho;Mohanty Swomitra K.;Frazier A. Bruno
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.5 no.1
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    • pp.1-10
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    • 2005
  • This paper presents the development of a microsystem for whole blood purification and electrophysiological analysis of the purified cells. Magnetophoresis using continuous diamagnetic capture (DMC) was utilized for whole cell purification and electrical impedance spectroscopy (EIS) was utilized for electrophysiological analysis of the purified cells. The system was developed on silicon and plastic substrates utilizing conventional microfabrication technologies and plastic microfabrication technologies. Using the magnetophoretic microseparator, white blood cells were purified from a sample of whole blood. The experimental results of the DMC microseparator show that 89.7% of the red blood cells (RBCs) and 72.7% of the white blood cells (WBCs) could be continuously separated out from a whole blood using an external magnetic flux of 0.2 T. EIS was used as a downstream whole cell analysis tool to study the electrophysiological characteristics of purified cells. In this work, primary cultured bovine chromaffin cells and human red blood cells were characterized using EIS. Further analysis capabilities of the EIS were demonstrated by successfully obtaining unique impedance signatures for chromaffin cells based on the whole cell ion channel activity.

Magnetophoretic Microseparators for Separating Blood Cells Based on Their Native Magnetic Properties (혈액 세포의 고유자성을 이용한 마이크로 자기영동 세포분리기)

  • Jung, Jin-Hee;Han, Ki-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.32 no.11
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    • pp.856-862
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    • 2008
  • This paper presents the characterization of a continuous magnetophoretic microseparator for separating white and red blood cells from peripheral whole blood cells based on their native magnetic properties. The magnetophoretic microseparator separated the blood cells using a high gradient magnetic separation (HGMS) method without the use of additives such as magnetic beads or probing materials. Experimental results show that the paramagnetic capture mode microseparator can continuously separate out 93.5% of red blood cells and 97.4% of white blood cells from diluted whole blood, and the diamagnetic capture mode microseparator can continuously separate out 89.7% of red blood cells and 72.7 % of white blood cells by using applying an external magnetic flux of 0.2 T using a permanent magnet.

Measurement of red cell deformability and whole blood viscosity using laser-diffraction slit rheometer

  • Sehyun Shin;Yunhee Ku;Park, Myung-Su;Suh, Jang-Soo
    • Korea-Australia Rheology Journal
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    • v.16 no.2
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    • pp.85-90
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    • 2004
  • The present study investigated the deformability of red blood cells (RBC) and its effect on whole blood viscosity using a laser-diffraction slit-rheometer (LDSR). The LDSR has been recently developed with significant advances in laser-diffractometry design, operation and data analysis. While shear stress levels in a slit flow are continuously decreasing, both the deformation of red blood cells and the shear stress were simultaneously measured. Additionally, the viscosity of whole blood was measured using the LDSR. The present study found that the whole blood viscosity is strongly dependent on the RBC deformability. The less deformable the RBCs are, the higher the blood viscosity is.

Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation

  • Wang, Jiying;Wang, Yanping;Wang, Huaizhong;Hao, Xiaojing;Wu, Ying;Guo, Jianfeng
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.471-478
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    • 2014
  • Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

A standardized method to study immune responses using porcine whole blood

  • Sameer-ul-Salam Mattoo;Ram Prasad Aganja;Seung-Chai Kim;Chang-Gi Jeong;Salik Nazki;Amina Khatun;Won-Il Kim;Sang-Myeong Lee
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.11.1-11.14
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    • 2023
  • Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ+CD3+ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ+ CD4-CD8+ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

Microdevice for Separation of Circulating Tumor Cells Using Embedded Magnetophoresis with V-shaped Ni-Co Nanowires and Immuno-nanomagnetic Beads

  • Park, Jeong Won;Lee, Nae-Rym;Cho, Sung Mok;Jung, Moon Youn;Ihm, Chunhwa;Lee, Dae-Sik
    • ETRI Journal
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    • v.37 no.2
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    • pp.233-240
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    • 2015
  • The novelty of this study resides in a 6"-wafer-level microfabrication protocol for a microdevice with a fluidic control system for the separation of circulating tumor cells (CTCs) from human whole blood cells. The microdevice utilizes a lateral magnetophoresis method based on immunomagnetic nanobeads with anti-epithelial cell adhesive molecule antibodies that selectively bind to epithelial cancer cells. The device consists of a top polydimethylsiloxane substrate for microfluidic control and a bottom substrate for lateral magnetophoretic force generation with embedded v-shaped soft magnetic microwires. The microdevice can isolate about 93% of the spiked cancer cells (MCF-7, a breast cancer cell line) at a flow rate of 40/100 mL/min with respect to a whole human blood/buffer solution. For all isolation, it takes only 10 min to process 400 mL of whole human blood. The fabrication method is sufficiently simple and easy, allowing the microdevice to be a mass-producible clinical tool for cancer diagnosis, prognosis, and personalized medicine.

Change in Rouleau Formation of Red Blood Cells by Pulse Magnetic Field Stimulus in the Hand (펄스자기장 자극에 의한 손의 적혈구 형태학적 변화)

  • Hwang, Do Guwn
    • Journal of the Korean Magnetics Society
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    • v.24 no.1
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    • pp.28-33
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    • 2014
  • The change of rouleau formation of red blood cells in the hand stimulated by pulse magnetic field having a maximum intensity of 0.27 Tesla and pulse duration of 0.102 msec was investigated. Before pulse magnetic field stimulus, the red blood cells of test subject were adjoined over ten and the flow of cells was slowed. However after the stimulus in the hand during 10 minutes, the red blood cells adjoined over tens was spreaded out each other and its motion was fast. Also the red blood cells of left hand unstimulated by pulse magnetic field were spreaded out each other, even though the right hand was stimulated during 10 minute. It prove that the rouleau formation of red blood cells can be improve in the whole body in spite of stimulus in the hand because the blood is flowing a whole body.

Reducing the Disposal of Unused Blood in the Operating Room (수술장에서의 혈액 폐기율 감소)

  • Cho, Moon Su;Lee, Yeoung Sook;Yu, Il Me
    • Quality Improvement in Health Care
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    • v.3 no.2
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    • pp.70-85
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    • 1997
  • Background : 6.1% of red blood cells and whole blood issued to the operating room was not transfused to the patients and discarded in Seoul National University Hospital in 1994. Objectives : We planned to set up an effective management program of blood in the operating room and we investigated whether this program could reduce the disposal rate of blood. Methods : We made a guideline of blood management in the operating room through a workshop. The guideline was revised after a preliminary application. The revised guideline was applied for 5 months from May to September in 1996. The disposal rate was compared before and after the installation of the new program. Results : 5,336 units of blood were issued to the operating room for 5 months. Disposal rate of red blood cells and whole blood was markedly reduced from 6.2% in May to 2.1% in September(p<0.05). The average disposal rate was 3.7% during the five months. Conclusion : We were able to reduce the disposal of unused blood in the operating room through the development and the application of a new blood management program.

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Screening of Anti-Atopic Dermatitis Material by Using NC/Nga Mouse Whole Blood System (NC/Nga 마우스 전혈을 이용한 항 아토피 피부염 물질 탐색)

  • Park, Dong-Hoon;Kim, Youn-Uck
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.98-105
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    • 2008
  • Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.

Morphological Review of Red Blood Cells After X-ray Irradiation (방사선 조사 후 적혈구의 형태적 고찰)

  • Tae-Jeong Ji
    • Journal of the Korean Society of Radiology
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    • v.17 no.2
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    • pp.223-230
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    • 2023
  • This study attempted to concider the morphological change of red blood cells after whole body irradiation. Blood samples used red blood cells of white mice and mouse after irradiation. Transmission electron microscope observation results, Anisocytosis was observed in red blood cells 20 days after 5 Gy irradiation. Triangle and tetrapod were observed for small red blood cell types. Poikilocytosis, sickle-shaped Drepanocyte, and Acantocyte were observed in general-sized red blood cells. Schizocyte was observed in red blood cells 20 days after 7 Gy irradiation. Scanning electron microscope observation results, Dacryocyte was observed with microcytes. It was also confirmed that red blood cells were get tangled with each other. In addition, polygonal shapes and half-moon shapes were also observed. In conclusion, it is judged that the modified form of pathological study is more important than the numerical change in the study of red blood cells by radiation exposure. In conclusion, it was confirmed that modified morphological studies are more important than numerical changes in the study of red blood cells by radiation exposure.