• Title/Summary/Keyword: total cell number

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Effects of Knockout Serum Replacement in the Culture Medium on the Proliferation of Porcine Fetal Fibroblasts In Vitro

  • Kim, Eun-Ju;Park, Jung-Joo;Choi, Young-Ju;Park, Sang Kyu;Roh, Sang-Ho
    • International Journal of Oral Biology
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    • v.35 no.1
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    • pp.1-5
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    • 2010
  • Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

Cryopreservation of Embryo by Concentration of Ethylene Glycol and Day 6, 7, 8, 9 Embryo in Korean Native Cattle (Hanwoo)

  • Park, Joung-Jun;Yoo, Han-Jun;Choi, Hye-Won;Cheong, Ki-Soo;Kim, Ji-Tae;Park, Choon-Keun;Yang, Boh-Suk
    • Journal of Embryo Transfer
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    • v.27 no.4
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    • pp.253-258
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    • 2012
  • This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozen-thawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h ($94.2{\pm}2.6%$, $94.67{\pm}3.4$ and $129.67{\pm}5.5$). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to $98.3{\pm}1.7%$ after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.

Modulation of Immune Parameters by Aging Process (노화에 따른 면역지표의 변화에 관한 연구)

  • Lee, Ji-Hye;Jung, Ji-Hye;Kim, Hyun-Sook
    • Journal of Nutrition and Health
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    • v.43 no.2
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    • pp.152-160
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    • 2010
  • The purpose of this study was to investigate the effects of aging process on the immunity in human subjects. In this investigation, nineteen families of three generations (daughters on college age, their mothers, and grandmothers) participated to avoid genetic variation among individuals. Dietary food records, anthropometric measurements and biochemical assessments of serum nutrients were used to evaluate the nutritional status of subjects. The immune parameters of subjects were assessed by the total and differential WBC count. Total B and T lymphocytes, and T cell subsets were quantified by flowcytometer. Serum immunoglobulin G, A, M concentrations were also measured as an index of humoral immunity. The result of this study can be summarized as follows: 1. Along with the aging process, body fat was found to be increased whereas lean body mass and total body water were diminished. Since there were no significant difference in serum vitamin E levels in all age groups, serum retinal concentrations tended to decrease as one gets old. 2. Although total number of T lymphocytes seemed to be unchanged, B lymphocytes and NK cell numbers were increased by aging. The Percentage of CD8 + lymphocytes was lower in the elderly subjects compared with the younger, resulting in higher ratio of CD4 +/CD8 + lymphocytes in the elderly. Serum Ig G and Ig A levels remained unchanged, but IgM levels were significantly decreased as the age processes continue. Taking all together, it could be suggested that the alteration of immune cell population by aging is selective and possibly nonage factors such as nutrition may be attributable to the change of immunity in the elderly. The nutritional status and aging process may selectively affect both the cell-mediated (CD8 +, CD4 + CD8 + ratio, NK cell) and humoral (B lymphocyte, Immunoglobulin M, G) immune parameters in human subjects.

Studies on Kagamboatang(KGBT) on the Hematopoiesis and Proliferation of Immune Function in Mice (가감보아탕(加減補兒湯)의 조혈(造血) 및 면역증진(免疫增進)에 관한 연구(硏究))

  • Kim Yun-Hee;Yoo Dong-Youl
    • The Journal of Pediatrics of Korean Medicine
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    • v.14 no.1
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    • pp.79-116
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    • 2000
  • The KGBT has been used to weak children with anorexia, fatigue, and growth retardation. This study was carried out to prove the effects of the hematopoiesis and the immune proliferation by KGBT. Previously, C57BL/6 mice was treated with cyclophosphamide(100mg/kg) for leukopenia, and then administered KGBT (concentration is 1.37 g/kg, 504 mg/kg, and 137 mg/kg) to the treated mice. The mice was analyzed expression of thrombopoietin(TPO), stem cell factor(SCF) and interleukin-3 from bone marrow cell, interleukin-10 (IL-10), and interferon-$ {\gamma}$(INF-${\gamma}$) from splenic cell, and NOSⅡ gene from macrophage using by RT-PCR. Also proliferation of immune cell was analyzed using 3H-thymidine uptake and flow cytometery in splenic cells. The results were obtained as follows ; 1. The total number of WBC, RBC and PLT was increased in the KGBT treated group than in the control group. 2. In vitro, the proliferation of splenic cells was increased in normal, control, and KGBT treated group. And Administration of KGBT was reduced the cytotoxicity by CTX. 3. In bone marrow cell, the gene expression of immune regulatory factor that associated with hematopoiesis, such as TPO, SCF, and IL-13 was increased in the KGBT treated group than control. 4 The titer of hemagglutinin and hemolysin was increased in the KGBT treated group than control. 5. In analysis of positive leucocytes from splenic cell of BALB/c mice, the subpopulation percent of CD4+, CD8+,and CD19+ was increased in the KGBT treated group than control. The KGBT has been used to weak children with anorexia, fatigue, and growth retardation. This study was carried out to prove the effects of the hematopoiesis and the immune proliferation by KGBT. Previously, C57BL/6 mice was treated with cyclophosphamide(100mg/kg) for leukopenia, and then administered KGBT (concentration is 1.37 g/kg, 504 mg/kg, and 137 mg/kg) to the treated mice. The mice was analyzed expression of thrombopoietin(TPO), stem cell factor(SCF) and interleukin-3 from bone marrow cell, interleukin-10 (IL-10), and interferon-$ {\gamma}$(INF-${\gamma}$) from splenic cell, and NOSⅡ gene from macrophage using by RT-PCR. Also proliferation of immune cell was analyzed using 3H-thymidine uptake and flow cytometery in splenic cells. The results were obtained as follows ; 1. The total number of WBC, RBC and PLT was increased in the KGBT treated group than in the control group. 2. In vitro, the proliferation of splenic cells was increased in normal, control, and KGBT treated group. And Administration of KGBT was reduced the cytotoxicity by CTX. 3. In bone marrow cell, the gene expression of immune regulatory factor that associated with hematopoiesis, such as TPO, SCF, and IL-13 was increased in the KGBT treated group than control. 4 The titer of hemagglutinin and hemolysin was increased in the KGBT treated group than control. 5. In analysis of positive leucocytes from splenic cell of BALB/c mice, the subpopulation percent of CD4+, CD8+,and CD19+ was increased in the KGBT treated group than control. 6. The expression of IL-10 gene was reduced in the KGBT treated group than control, whereas the expression of INF-${\gamma}$ was increased in the KGBT treated group. 7. In macrophage, the production of NO and gene expression of NOSH was increased in the KGBT treated group than control. 8. After infection of EMC virus, the survival time of infected mice was longer in the KGBT treated group than control.

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A STUDY ON OSTEOBLAST-LIKE CELL RESPONSES TO SURFACE-MODIFIED TITANIUM

  • Hong Min-Ah;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Su;Lee Jae-Il
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.3
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    • pp.300-318
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    • 2003
  • Statement of problem: The success of implants depends on intimate and direct contact of implant material on bone tissue and on functional relationship with soft tissue contact. Creation and maintenance of osseointegration depend on the understanding of the tissue's healing, repairing, and remodeling capacity and these capacities rely on cellular behavior. Altering the surface properties can modify cellular responses such as cell adhesion, cell motility, bone deposition, Therefore, various implant surface treatment methods are being developed for the improved bone cell responses. Purpose: The purpose of this study was to evaluate the responses of osteoblast-like cells to surface-modified titanium. Materials and Methods: The experiment was composed of four groups. Group 1 represented the electropolished surface. Group 2 surfaces were machined surface. Group 3 and Group 4 were anodized surfaces. Group 3 had low roughness and Group 4 had high roughness. Physicochemical properties and microstructures of the discs were examined and the responses of osteoblast-like cells to the discs were investigated. The microtopography was observed by SEM. The roughness was measured by three-dimension roughness measuring system. The microstructure was analyzed by XRD, AES. To evaluate cell responses to modified titanium surfaces, osteoblasts isolated from calvaria of neonatal rat were cultured. Cell count, morphology, total protein measurement and alkaline phosphatase activities of the cultures were examined. Results and Conclusion: The results were as follows 1. The four groups showed specific microtopography respectively. Anodized group showed grain structure with micropores. 2. Surface roughness values were, from the lowest to the highest, electropolished group, machined group, low roughness anodized group, and high roughness anodized group. 3. Highly roughened anodized group was found to have increased surface oxide thickness and surface crystallinity. 4. The morphology of cells, flattened or spherical, were different from each other. In the electropolished group and machined group, the cells were almost flattened. In two anodized groups, some cells were spherical and other cells were flattened. And the 14 day culture cells of all of the groups were nearly flattened due to confluency. 5. The number of attached cells was highest in low roughness anodized group. And the machined group had significantly lower cell count than any other groups(P<.05). 6. Total protein contents showed no difference among groups. 7. The level of alkaline phosphatase activities was higher in the anodized groups than electropolished and machined groups(P<.05).

Iron Status of the Adolescent Females before and after Menarche (초경 전후 사춘기 여성의 철 영양에 관한 연구)

  • 임현숙;정은숙
    • Journal of Nutrition and Health
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    • v.36 no.6
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    • pp.646-652
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    • 2003
  • This study was performed to determine the iron status of the adolescent Korean girls before and after menarche. The 101 subjects aged 11-13 years who attending in an elementary school in Mokpo were recruited. They were divided into pre-menarche (A) group or post-menarche (B) group based on their menstruation status. The latter subjects were sub-divided into one of the four groups according to the times of their menstruation B-I( $\geq$ 3 times), B-II (4-6 times), B-III (7-9 times) or B-W ( $\geq$ 10 times). In the total subjects, dietary iron intake, 11.3 mg/day, was below the Korean RDA for iron, the percentage of heme iron to total iron intake, 15%, and the bioavailability of dietary iron, 12.3%, seemed to be low. And their body iron storage, 140.8 mg, seemed to be insufficient. However, they tended to meet body's iron requirement in the cell level. Red blood cell number (RBC), hematocrit (Hct), and hemoglobin (Hb) level in the total subjects were 4.5 1012/I, 39.3%, and 13.0 g/㎗, respectively. The subjects in B group had lower (p<0.05) RBC and Hct compared to those in A group and the prevalence of iron-deficiency anemia tended to be high. Serum iron, ferritin, and soluble transferrin receptor (sTfR) and sTfR:ferritin ratio were 86.7 $\mu\textrm{g}$/d, 17.6 $\mu\textrm{g}$/l, 3.58 mg/1, and 230, respectively. Those four indices were not significantly different among the groups. The results of this study imply that, although there a tendency to affect negatively iron status, menstrual blood loss in adolescent females does not deteriorate obviously their iron status during the relatively short period up to 1 you. However, it should be better to improve their iron status after starting menarche by increasing iron intake, especially heme-iron, and enhancing factors for iron absorption.

Effect of splenectomy on the blood and marrow megakaryocyte picture in dogs (견(犬)에 있어서 비장적출(脾臟摘出)이 혈액(血液) 및 골수거대핵세포상(骨髓巨大核細胞像)에 미치는 영향(影響))

  • Hong, Kyung-tae;Lee, Hyun-beom;Lee, Keun-woo
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.327-336
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    • 1993
  • Present experiments were undertaken in order to clarify the effect of splenectomy on the hematology and marrow megakaryocyte picture and to know the genesis of postsplenectomy thrombocytosis in dogs. Six mongrel dogs weighing 8.5~18㎏ were used, of which three were splenectomized and the other three were laparotomized for comparison. Erythrocyte count, total and differential leukocyte counts, thrombocyte count and packed cell volume measurement were made using the blood samples. In addition, bone marrow samples obtained from the femur at 7th and 23rd day of the operation were examined for the number per low-power field, the diameter, and the distribution frequency of the megakaryocyte. From these experiments, following results were obtained : Erythrocyte count and packed cell volume showed significant decrease beginning on the 15th day of splenectomy. Total and differential leukocyte counts showed marked increase for the first 2 days of postsplenectomy. The thrombocyte count of splenectomized dogs increased from the 2nd day of the operation, reached to the peak count on the 15th day, and returned to the preoperation count by the 28th day. The megakaryocyte count per low-power field of the biopsied preparation increased in according to the increase in thrombocyte count. The megakaryocyte diameter of splenectomized dog showed no increase on the 7th or 23rd day of the operation. However, the distribution frequency of the larger megakaryocyte was higher in the splenectomized dogs than in the laparotomized dogs. The total plasma protein concentration showed no significant change after splenectomy or laparotomy. From these results, it may be concluded that the postsplenectomy thrombocytosis results from the increased megakaryocytopoesis or the activated thrombocytopoesis of the marrow megakaryocytes.

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A Study on the Nutritional Status of Children Living in Rural Area and City Orphan Asylum -II. Hematological view- (농어촌(農漁村)과 시설거주(施設居住) 아동(兒童)의 영양상태(營養狀態)에 관(關)한 연구(硏究) -제2보(第2報) : 혈액성장(血液性狀)을 중심(中心)으로-)

  • Park, Bock-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.108-116
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    • 1985
  • This study was designed to investigate the nutritional anemic state and hematological findings of children, aged 11 and 12 years, living in Haenam Koon (living at home) and in Mokpo City (living in an orphan asylum) located in Chonnam. The mean red blood cell number of male was higher than female in both groups of living in the rural area and in the city orphan asylum (p<0.01), but the white blood cell count was not significantly different. The levels of average hemoglobin, hematocrit and mean corpuscular hemoglobin concentration of the children in both groups were similar, and 19.8% of children living at home in the rural area and 32.1% of children living in the city orphan asylum were anemic. The mean levels of serum total protein, albumin and A/G ratio in children of both group were not statistically different, and 18.4% of children living at home in the rural area and 13.2% of children living in the city orphan asylum were insufficient in the serum total protein value. The average serum cholesterol level of children living at home in the rural area was higher than that of children living in the city orphan asylum(p<0.01), and that of female living at home was higher than that of male (p<0.05). The mean levels of serum iron, total iron binding capacity and transferrin saturation of children living at home in the rural area were significantly higher than those of children living in the city orphan asylum (p<0.01).

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Shear lag effect of varied sectional cantilever box girder with multiple cells

  • Guo, Zengwei;Liu, Xinliang;Li, Longjing
    • Structural Engineering and Mechanics
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    • v.84 no.3
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    • pp.295-310
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    • 2022
  • This paper proposes a modified bar simulation method for analyzing the shear lag effect of variable sectional box girder with multiple cells. This theoretical method formulates the equivalent area of stiffening bars and the allocation proportion of shear flows in webs, and re-derives the governing differential equations of bar simulation method. The feasibility of the proposed method is verified by the model test and finite element (FE) analysis of a simply supported multi-cell box girder with constant depth. Subsequently, parametric analysis is conducted to explore the mechanism of shear lag effect of varied sectional cantilever box girder with multiple cells. Results show that the shear lag behavior of variable box-section cantilever box girder is weaker than that of box girder with constant section. It is recommended to make the gradient of shear flow in the web with respect to span length vary as smoothly as possible for eliminating the shear lag effect of box girder. An effective countermeasure for diminishing shear lag effect is to increase the number of box chambers or change the variation manner of bridge depth. The shear lag effect of varied sectional cantilever box girder will get more server when the length of central flanges is shorter than 0.26 or longer than 0.36 times of total width of top flange, as well as the cantilever length exceeds 0.29 times of total length of box's flange. Therefore, the distance between central webs can adjust the shear lag effect of box girder. Especially, the width ratio of cantilever plate with respect to total length of top flange is proposed to be no more 1/3.

The Effects of Bogimakseong-bang(補氣膜性方) Treatment on cBSA-induced Membranous Nephropathy in Mouse Model (보기막성방(補氣膜性方)이 Cationized Bovine Serum Albumin투여로 유발된 Mouse의 Membranous Nephropathy에 미치는 영향)

  • Lee, Jung-Won;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.4
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    • pp.1083-1099
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    • 2008
  • Objective : We aimed to identify the effects of Bogimakseong-bang(BGMSB) treatment on cBSA-induced MN in a mouse model. Methods : We divided 20 mice into 4 groups. The normal group (NR) had no treatment. We used cBSA to induced MN to the other 3 groups. One group (CT) was treated with cBSA (7mg/kg i.p) only. The second (BG-250) was treated with cBSA (7mg/kg i.p) and BGMSB extract (250mg/kg, p.o). The third group (BG-500) was treated with cBSA(7mg/kg i.p) and BGMSB extract (500mg/kg, p.o). After cBSA and BGMSB extract treatment for 4 weeks, proteinuria, serum albumin, total cholesterol, serum creatinine, BUN, total nucleated spleen cell number and total infiltrated kidney cell number of all groups were measured. CD3e+/CD19+ and CD4+/CD8 cells ratio of peripheral blood, kidney and spleen of all groups were analyzed. $IL-1{\beta}$ and $TNF-{\alpha}$, IL-6, IgG, IgM, and $IFN-{\gamma}$ levels of all groups were gauged. Histological analysis of kidney tissue and immunohistochemical staining (CD4 CD8) of kidneys were observed. Results : Proteinuria significantly decreased and serum albumin increased in groups treated with cBSA and BGMSB extract compared with the control. Total cholesterol decreased but not significantly. CD3e+/CD19cells ratio of peripheral blood decreased. CD3e+/CD19+ and CD4+/CD8 cells percentage of kidney and spleen showed no significant change. Level of $IL-1{\beta}$, $TNF-{\alpha}$ and IL-6 significantly decreased. and $IFN-{\gamma}$ increased but has not significantly. Concentration of IgG and IgM significantly decreased compared with control. Thickness of GBM decreased on histological analysis of kidney. Deposition of CD4 and CD8 decreased on immunohistochemical staining of kidney. Conclusions : According to the above result, BGMSB had a significant effect for treating MN which is cBSA-induced.

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