• Title/Summary/Keyword: Hemocytometer

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Antiproliferative Effect of Mistletoe Extract Added Kimchi in Human Lung Carcinoma A549 Cells (겨우살이 물추출물 첨가 김치의 A549 인체 폐암 세포 증식저해 효과)

  • Kil, Jung-Ha
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1507-1514
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    • 2017
  • The purpose of this study aimed at examining the antiproliferative effect of kimchi (kimchi B) adding mistletoe extract known as an anticancer function to improve the functions of kimchi. The study investigated the antiproliferative effect through hemocytometer counts and MTT assay, apoptosis induction through DAPI staining, and mRNA expression through RT-PCR using human lung carcinoma A549 cells. The standardized kimchi (Kimchi A) was used as a control group. As a result of hemocytometer counts and the MTT assay, it was found that kimchi samples inhibited the growth of A549 cells in a concentration-dependent manner. Kimchi B induced apoptosis in A549 cells through DAPI staining. The apoptosis induced by kimchi B was associated with the increase in the expression of pro-apoptotic Bax and with the decrease in the expression of anti-apoptotic Bcl-2 and Bcl-xL. Also, kimchi B influenced the increase in the expression of p21 mRNA, but did not have the effect on the expression of p53 mRNA. In conclusion, the antiproliferative effect of kimchi B was due to apoptosis induced by increasing Bax and decreasing Bcl-2, and increasing p21. The findings will be utilized to develop kimchi with the improved function for the patients having cancer.

AN EXPERIMENTAL STUDY ON THE CHANGE OF THE RADIOSENSITIVITY OF SEVERAL TUMOR CELL LINES AND PRIMARY CULTURED GINGIVAL FIBROBLAST (수종의 암세포주와 치은섬유아세포에서 방사선의 양과 분할조사에 따른 세포활성도와 독성의 변화에 관한 연구)

  • Lee Sam-Sun;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.107-122
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    • 1997
  • Radiation sensitivity data was generated for two human cancer cell lines(KB, RPMI 2650) and human primary gingival fibroblast was tested three times using a viable cell number counting with a hemocytometer, MTT(3-[4,5-Dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay, and LDH(Lactate dehydrogenase) assay. Single irradiation of 2, 4, 6, 10, 15, 20Gy were applied to the tumor cell lines and the primary cultured gingival fibroblast The two fractions of 4Gy and 10Gy were seperated with a 4 hour time interval. The irradiation was done with 241.5cGy/min dose rate using /sup 137/Cs MK cell irradiator at room temperature. The obtained results were as followed : 1. There was significantly different viable cell numbers as the amount of radiation dose on the tested cells were cell number counted with a hemocytometer. In fractions, there were more viable cells remaining. 2. Phase-contrast microscopically, radiation-induced morphologic changes were pronounced on the tumor cells, however, almost no differences on the gingival fibroblast. 3. There was significantly different absorbance at 2Gy on RPMI 2600, 4Gy on KB and GF in MTT assay. In fractions, the absorbance was significantly higher on KB. 4. The level of extracellular LDH activity in the experimental group was significantly higher in the 2-4Gy than the control group. 5. The total level of extracellular and intracellular LDH activity was decreased as increased amounts of radiation dose was applied.

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Fabrication and Application of Micro Polymer Chip Platform for Rare Cell Sample Preparation (희귀 세포 샘플 준비를 위한 마이크로 폴리머 칩 플랫폼 제작 및 활용)

  • Park, Taehyun
    • Journal of the Korea Convergence Society
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    • v.9 no.3
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    • pp.217-222
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    • 2018
  • In this paper, a new micro polymer chip platform and protocol were developed for rare cell sample preparation. The proposed platform and protocol overcome the current limitation of the dilution method which is based on statistics and the FACS method which expensive and requires fluorescence staining. It allows collecting exact number of target cells simply and selectively because the cells are visually confirmed during the collecting process. The collected cells can be transported or spiked into a desired locations, such as a microchamber, without cell loss. This research may applicable not only to a rare cell sample preparation for Lab on a Chip cancer diagnosis, but also to a single/double/multiple cell sample preparation for a cell analysis field. To verify this platform and protocol, five human breast cancer cells (MCF-7) were collected and transported into a hemocytometer chamber.

Analysis of Blood Cell Images Using Smartphone-based Mobile SmartScope (스마트폰 기반 Mobile SmartScope를 이용한 혈구 영상 분석)

  • Park, Choonho;Cho, Myoung-Ock;Lee, Donghee;Kim, Jung Kyung
    • Journal of the Korean Society of Visualization
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    • v.10 no.2
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    • pp.25-31
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    • 2012
  • High-performance smartphones, equipped with a digital camera and an application software, can render conventional bench-top laboratory instruments mobile at affordable costs. As the smartphone-based devices are portable and wireless, they have wide applications especially in providing point-of-care (POC) tests in resource-constrained areas. We developed a hand-held diagnostic system, Mobile SmartScope, which consists of a small optical unit integrated with a smartphone. The performance of the SmartScope was favorably compared with that of conventional light microscopy in detecting and quantifying red blood cells. We also evaluated the fluorescence detection limit of the SmartScope incorporated with a blue light-emitting diode and appropriate optical filters by using fluorescently labeled microbeads for intensity calibration.

A Novel Cell Count Method Using Micro Lattice Engraved On A Culture Dish (미세 격자가 형성된 배양 접시를 이용한 새로운 세포 계수 방법)

  • Son, Sang-Uk;Choi, Yo-Han;Lee, Seung-Seob
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.28 no.8 s.227
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    • pp.1183-1189
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    • 2004
  • A novel cell count method, which can improve the count efficiency and reduce contamination problem, was presented using micro lattice engraved on culture dish. The micro lattice has feature of $50{\mu}m{\times}50{\mu}m$ rectangular shape, $2{\mu}m$ line width, and $2{\mu}m$ depth in $3mm{\times}3mm$ area. In this paper, nickel mold was fabricated with thickness of 3mm and diameter of 80 mm, and transcription of the micro lattice on a polystyrene cell culture dish was performed by hot embossing at $200\;^{\circ}C$. The tedious and error-prone harvest/load processes of conventional cell counts with a hemocytometer could be omitted, and these advantages became magnified during periodical counts involving long-term cultures. SupT1 cells and HeLa cells were cultivated with the dish for 7 days in $CO_2$ incubator and counted as $371.84/mm^2$ and $123.36/mm^2$, respectively, during the cultivation without harmful effects on the cells.

Study on Improved Diluting Fluids for Direct Counts of Total Leukocyte, Eosinophil and Other Leukocytes (총백혈구수(總白血球數), 호산구수(好酸球數) 및 기타 백혈구수(白血球數)의 직접계산(直接計算)을 위한 개량희석액(改良稀釋液)에 관한 연구(硏究))

  • Lee, Bang Whan
    • Korean Journal of Veterinary Research
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    • v.16 no.2
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    • pp.105-114
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    • 1976
  • The study was conducted for an attempt to improve the diluting fluid for total leukocyte count, and to prepare a multipurpose diluting fluid for concurrent direct counts of total leukocytes, eosinophils and the other leukocytes. Through the experiment, two better fluids for total leukocyte count of blood of human, bovine, swine, canine and rabbit were selected, and which conserved cell morphology of leukocytes better than $T{\ddot{u}}rk$-solution. Each formula of two fluids were given as under R I and R II. Formula of multipurpose diluting fluid selected in the experiment was given as under III. With this fluid, direct counts of total leukocytes, eosinophils and probably basophils of blood of human, bovine and swine were practicable concurrently in the same counting chamber of a hemocytometer. In this fluid, eosinophils were stained red in the part of eosinophilic granules and blue in other part of cell, and basophils were stained dark blue like as a lump of black granules, staining three other leukocytes faint blue. Eosinophils of canine blood were not so enough red those in other animal and human and eosinophils of rabbit blood were not distinguishable from pseudoeosinophils in this fluid.

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Apoptosis of Human Lung Carcinoma Cells through the Inhibition of Bcl-2 Expression and Activation of Caspase by Chungjogupae-tang (인체폐암세포에서 Bcl-2 발현저하 및 caspase 활성을 통한 청조구폐탕의 apoptosis 유발에 관한 연구)

  • Cho, In-Joo;Gam, Chul-Woo;Kim, Ki-Tak;Park, Dong-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.93-97
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    • 2007
  • We previously reported the anti-proliferative effect of Chungjogupae-tang (CJGPT) in human lung carcinoma A549 cells, which was associated with the induction of cyclin-dependent kinase inhibitor p21 in a tumor suppressor p53-independent manner. CJGPT treatment also resulted in the inhibition of prostaglandin E2 release A549 cells by the down-regulation of cyclooxygenase-2. In the present study, we investigated the pathway of the induction of apoptotic cell death by CJGPT in A549 cells. It was found that CJGPT could inhibit the cell viability and induce the apoptotic cell death of A549 cells in a dose-dependent manner as measured by hemocytometer counts, flow cytometry analysis and agarose gel electrophoresis. Apoptosis of A549 cells by CJGPT was associated with a down-regulation of anti-apoptotic Bcl-2 and inhibitor of apoptosis proteins (IAPs) expression. Additionally, DNA fragmentation by CJGPT was connected with the activation of inhibitor of caspase-activated DNase/DNA fragmentation factor 45 (ICAD/DFF45) protein expression.

Induction of Apoptotic Cell Death by Healthful Decoction Utilizing Phellinus Linteus in Human Lung Carcinoma Cells (상황을 이용한 보건기능 개선제의 인체폐암세포 apoptosis 유발에 관한 연구)

  • Park Cheol;Lee Yong Tae;Kang Kyung Hwa;choi Byung Tae;Jeong Young Kee;Choi Yung Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.759-766
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    • 2004
  • In the present study, we investigated the effects of aqueous extract of the healthful decoction utilizing Phellinus linteus (HDPL) on the cell growth of human lung carcinoma tumor cell line A549. Exposure of A549 cells to HDPL resulted in growth inhibition and induction of apoptosis in a dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometric analysis. This increase in apoptosis was associated with inhibition and/or degradation of apoptotic target proteins such as poly(ADP-ribose) polymerase (PARP), b-catenin and phospholipase C- 1 (PLC- 1) protein. HDPL treatment induced the down-regulation of anti-apoptotic Bcl-2 expression, an anti-apoptotic gene, however, the level of Bax. a pro-apoptotic gene, was increased by HDPL treatment. In addition, HDPL-induced apoptotis of A549 cells was connected with activation of caspase-3 and caspase-9 protease in a dose-dependent manner, however, the levels of inhibitor of apoptosis proteins family were remained unchanged. Taken together, these results indicated that the anti-proliferative effects of HDPL were associated with the induction of apoptotic cell death through regulation of several major growth regulatory gene products such as Bcl-2 family expression and caspase protease activity, and HDPL may have therapeutic potential in human lung cancer.

Attachment of Human Gingival Fibroblasts to Commercially Pure Titanium Surfaces with Different Instruments;A comparative Study in Vitro (기구조작에 따른 순수 타이타늄 표면 변화와 치은 섬유아 세포 부착에 관한 연구)

  • Seo, Sung-Chan;Song, In-Taeck;Lim, Jeong-Su;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.29 no.3
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    • pp.607-621
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    • 1999
  • This study examined the human fibroblasts cell attachment to commercially pure titanium surface which had been instrumented by 3 types of periodontal instruments. Commercially pure titanium plates were uniformly scaled using plastic, stainless steel, titanium curette. these all experimental groups 65 undirectional strokes with the designated curettes. Alteration of the surfaces due to instrumentation was evaluated by Form Talysurf(R) and reported as Ra value(mean surface roughness). Then other experimental groups were immersed in a cell suspension of human gingival fibroblasts($1{\times}10^5$ cell/ml). After 3 days of culture, cell attachment and morphology was observed by SEM, and attached cell were counted by Hemocytometer. A significant difference in mean Ra value was observed for surface instrumented by metal curette compared to either control surface or surface instrumented by the plastic curette(P<0.01). No stastically significant difference was noted between control surface and those instrumented by the plastic curette. SEM observation showed that cell morphology and attachment to the commercially pure titanium plate was similar appearance on the all experimental groups. Experimental groups instrumented by titanium curette and stainless steel curette were more attached cell number than control group, but experimental group instrumented by plastic curette were similar with control groups(P<0.01). In summary, metal curette produced an significant alteration of the commercially pure titanium surface and more favorable surface topography for cell attachment. Otherwise plastic curette was insignificantly altered the commercially pure titanium surface(P<0.01).

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Goat Milk Yoghurt by Using Lacto-B Culture Modulates the Production of Tumor Necrosis Factor-Alpha and Interleukin-10 in Malnourished Rats

  • Nurliyani, Nurliyani;Kandarina, B.J. Istiti;Kusuma, Sari;Trisnasari, Yunita Dewi
    • Food Science of Animal Resources
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    • v.34 no.1
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    • pp.88-98
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    • 2014
  • Total spleen lymphocytes, lymphocyte proliferation, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and interleukin-10 (IL-10) in spleen lymphocyte culture were studied in malnourished Wistar rats fed with goat milk yoghurt. Malnourished rats were created by using standard feed restriction as much as 50% of normal rats for 21 d. Goat milk yoghurt containing three types of microorganism e.g., Lactobacillus acidophilus, Sterptococcus thermophilus and Bifidobacterium longum derived from Lacto-B culture in powder form. After 21 d, the rats continued to receive restricted feeding and supplemented with goat milk yoghurt for 7 d. Total splenocytes were counted by hemocytometer. Splenocytes proliferation was expressed as stimulation index, whereas the TNF-${\alpha}$ and IL-10 of spleen lymphocyte culture were measured by ELISA technique. The total number of splenocytes and stimulation index of splenocytes in moderate malnourished and normal rats supplemented with goat milk yoghurt was not significantly different. The level of TNF-${\alpha}$ in the rat supplemented with goat milk yoghurt was lower (p<0.05) than the control group, whereas the level of IL-10 in the rat supplemented with goat milk yoghurt was higher (p<0.05) than the control group. In conclusion, goat milk yoghurt supplementation in malnourished rats could decrease TNF-${\alpha}$ as a representation of the pro-inflammatory cytokine, while it increases IL-10 as a representation of the anti-inflammatory cytokine.