• The Journal of Advanced Prosthodontics
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• 제6권5호
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• pp.379-386
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• 2014

PURPOSE. These days, mesenchymal stem cells (MSCs) have received worldwide attention because of their potentiality in tissue engineering for implant dentistry. The purpose of this study was to evaluate various growth inducing factors in media for improvement of acquisition of bone marrow mesenchymal stem cells (BMMSCs) and colony forming unit-fibroblast (CFU-F). MATERIALS AND METHODS. The mouse BMMSCs were freshly obtained from female C3H mouse femur and tibia. The cells seeded at the density of $10^6$/dish in media supplemented with different density of fetal bovine serum (FBS), $1{\alpha}$, 25-dihydroxyvitamin (VD3) and recombinant human epidermal growth factor (rhEGF). After 14 days, CFU-F assay was conducted to analyze the cell attachment and proliferation, and moreover for VD3, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was additionally conducted. RESULTS. The cell proliferation was increased with the increase of FBS concentration (P<.05). The cell proliferation was highest at the density of 20 ng/mL rhEGF compared with 0 ng/mL and 200 ng/mL rhEGF (P<.05). For VD3, although the colony number was increased with the increase of its concentration, the difference was not statistically significant (P>.05). CONCLUTION. FBS played the main role in cell attachment and growth, and the growth factor like rhEGF played the additional effect. However, VD3 did not have much efficacy compare with the other two factors. Improvement of the conditions could be adopted to acquire more functional MSCs to apply into bony defect around implants easily.

• Journal of Nutrition and Health
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• 제29권8호
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• pp.889-898
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• 1996

인슐린 비의존형 당뇨병환자의 심리적 스트레스와 스트레스호르몬 및 영양상태와의 상과관계를 알아보고자 인슐린 비의존형 당뇨병환자 34명을 대상으로 설문지를 사용하여 직접면접을 통해 심리적 스트레스로 우울, 불안을 측정하였고 24시간 회상법에 의한 영양섭취상태, 에너지 소비량 등을 조사하였고, 공복시 혈액을 채취하여 일반혈액검사에 스트레스호르몬, 아미노산 등을 분석하였다. 우울과 불안은 CED-S와 STAI-S의 방법으로 측정하였고, 영양섭취상태는 전산분석하였다. 그리고 catecholamine, hemoglobin A1C와 아미노산은 HPLC로 분석하였으며 cortisol과 glucagon은 RIA방법으로 분석하였다. 본 연구의 결과, 당뇨병환자의 우울과 불안점수는 대조군에 비해 높았는데 특히 불안은 유의적으로 높았다. 당뇨병환자의 스트레스호르몬을 보면 총 catecholamine은 329.18$\pm$111.49pg/ml이었고 norepinephirne은 233.95$\pm$73.99pg/ml이었으며 epinephrine은 94.03$\pm$75.97pg/ml이었다. Cortisol은 13.18$\pm$5.55ul/ml이었으며 gulcagon은 171.50$\pm$62.50pg/ml이었다. 당뇨병환자의 총 catecholamine, norpinephrine, cortisol 그리고 glucagon 등은 대조군에 비해 유의적으로 높았으며 epinephrine은 높은 경향을 나타냈다. 당뇨병환자의 영양소섭취량은 대조군에 비해 차이가 없었다. 그러나 당뇨병환자의 경우 열량과 칼슘은 권장량보다 적었으나 다른 영양소들은 권장량보다 많이 섭취하였다. 공복혈당, 평상시 공복혈당 (3개월 평균 공복혈당) 그리고 hemoglobin A1C는 각각 184.18$\pm$74.22mg/dl, 177.76$\pm$42.77mg/dl와 8.84$\pm$2.82%이었다. 당뇨병환자의 혈장아미노산의 농도는 일반적으로 정상범위에 속하였다. 당뇨병환자의 불안은 norepinephrine과 평상시 공복혈당과 유의적인 양의 상관관계를 나타내었다. 또한 glucagon, 평상시 공복혈당과 hemoglobin A1C는 분지아미노산 (BCAA, leucine, isoleucine and valine)과 유의적인 양의 상관관계를 나타내었다.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.65-70
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• 2012

귀중한 한국재래닭의 배반엽세포를 냉동보존하고, 키메라 닭을 통한 재래종의 복원을 도모하는 방법을 실용화하기 위해서, 한국재래종의 배반엽세포에 있어 최적의 동결 방법에 대해 검토했다. 배반엽세포의 동결은 세포를 단리한 후, 항동해제와 소태아혈청(FBS)를 함유하고 있는 동결용액 중에 부유하고 수지성 동결용기 중에 넣고, $-7^{\circ}C$에서 동결시키고 나서 분당 $-1^{\circ}C$에서 $-35^{\circ}C$까지 냉각하고 액체질소 안에 침지시켜서 실험을 진행했다. 동결조작 중에서 (1) 동결 전의 세포의 단리 방법, (2) FBS 농도, (3)동결 시 세포밀도가 동결융해 후의 세포생존율에 미치는 영향을 조사했다. (1) 세포의 단리 방법을 피펫팅으로부터 시험관 믹서에 의한 단시간의 flushing으로 변경하면, 동결융해 후의 세포의 생존율이 29%부터 51%로 향상된다. (2) 동결용액 속 FBS 농도를 20%에서 80%로 증가시키면 동결융해 후의 생존율이 28%에서 35%로 증가한다. 또한, (3) 융해 후, 생존율은 (2개의 배자/0.5 ml) 처리군에서의 동결은 34%인 반면에, (20개의 배자/0.5 ml)에서는 44%였다. 더욱이, 이 세 가지 개선점을 조합함으로써 동결융해 후의 생존율은 60%로, 개선 전의 41%에 비하여 크게 개선이 될 수 있고, 한국재래종의 배반엽세포의 동결보존의 실용화가 보다 더 향상될 수 있는 방법이 될 수 있음을 시사한다.

• 한국식품영양과학회지
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• 제38권6호
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• pp.663-666
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• 2009

산수유 에탄올 추출물이 환경호르몬인 dioxin 및 bisphenol A(BPA)에 의해 유도된 인체 전립선 암세포인 RC58T/h/SA#4(prostate epithelial primary cancer cell lines)의 성장을 억제하는지를 조사하였다. 즉 산수유 에탄올 추출물을 10, 100, 300 및 $500{\mu}g/mL$ 농도로 RC58T/h/SA#4에 처리했을 시 $500{\mu}g/mL$ 농도 이상에서 높은 성장 억제율을 나타내었으며, dioxin과 bisphenol A를 다양한 농도로 처리한 결과 각각 1 nM, $0.1{\mu}M$ 농도에서 가장 높은 전립선암세포 증식을 유도하였다. 한편 환경호르몬(dioxin, BPA)에 의해 RC58T/h/SA#4의 증식을 유도시킨 후 산수유 에탄올 추출물을 10, 100, 300 및 $500{\mu}g/mL$ 농도로 처리한 결과 대조군에 비하여 산수유 에탄올 추출물 $500{\mu}g/mL$ 농도에서 그 억제율이 가장 높았다.

• 대한치과교정학회지
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• 제23권3호
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• pp.415-425
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• 1993

Fluoride is one of the most potent stimulators of bone formation in vivo. But its direct effects on osteoblast is not yet clear This study was to investigate the effects of Sodium fluoride on alkaline phosphatase(ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in Murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in $\alpha-Minimal$ essential medium $(\alpha-MEM)$ supplemente with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concentration of Sodium fluoride. The ALP activity was assayed by the method of Lowry with disodium phenyl phosphated as substrate. cAMP formation was measured by Radioimmuno Assay(RIA). Type I $\alpha$ 2 collagen ribonucleic acid(mRNA) expression was studied by Nothern blot analysis. The results were as follows: 1. cAMP level was increased by PTH in MC3T3-E1 cells. 2. Sodium fluoride showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. 3. Sodium fluoride increased ALP activity at cocentration of $2{\mu}M,\;4{\mu}M,\;and\;10{\mu}M$ significantly different from control at the 0.001 level. ALP activity revealed maximum value at $10{\mu}M$ in this study. 4. Nothern blot analysis of Sodium fluoride treated cells, using Type I $\alpha$ 2 collagen prove, revealed significant increase at $10{\mu}M$ in MC3T3-E1 cells.

• 대한지역사회영양학회지
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• 제25권5호
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• pp.396-405
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• 2020

Objectives: This study was conducted to compare the nutrient intake of normal healthy adults with those having hypercholesterolemia. Methods: We analyzed data from the 6th Korea National Health and Nutrition Examination Survey (KNHANES VI). A total of 12,636 adults (5,223 males and 7,413 females) aged 19 or older were included in the study. Results: Males with hypercholesterolemia were older and had a higher waist circumference, body mass index, fasting blood sugar levels (FBS) and serum triglyceride (TG) concentrations compared to the normal group. Females with hypercholesterolemia were older and had higher FBS levels and serum TG concentrations compared to the normal group. While comparing nutrient intake by the 24-hour recall method, the male normal group showed a higher intake of fat, saturated fatty acid (SFA), monounsaturated fatty acid (MUFA), vitamin A and thiamin compared to the hypercholesterolemic group. However, the male normal group had a lower intake of iron and vitamin C compared to the hypercholesterolemic group. The female normal group had a higher intake of energy, protein, fat, SFA, MUFA, polyunsaturated fatty acids, cholesterol, riboflavin, and niacin compared to the hypercholesterolemic group, but had a lower intake of iron compared to the hypercholesterolemic group. A comparison of nutrient intake by food frequency questionnaire (FFQ) showed the following: There was no significant difference in nutrient intake between the normal men and women and those with hypercholesterolemia. After adjustment for confounding factors, nutrient intake by FFQ of the male normal group showed higher levels of n-3 fatty acid and vitamin C compared to the group with hypercholesterolemia. However, there was no significant difference in nutrient intake between the two groups of women. Conclusions: The average intake of n-3 fatty acids and vitamin C of the male group with hypercholesterolemia was lower than that of the normal group. However, since KNHANES is a cross-sectional study, prospective cohort studies are required to analyze the risk factors of hypercholesterolemia.

• Animal Bioscience
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• 제36권11호
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• pp.1757-1768
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• 2023

Objective: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. Methods: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17β-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. Results: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17β-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. Conclusion: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

• International Journal of Oral Biology
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• 제37권2호
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• pp.43-50
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• 2012

The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we selected four types of novel small molecule candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

• Asian-Australasian Journal of Animal Sciences
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• 제19권1호
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• pp.119-125
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• 2006

The present study including two experiments was designed to determine the effect of media containing different rare earth elements (REE) on proliferation and fatty acids accumulation in 3T3-L1 cell cultures. In Experiment 1, 3T3-L1 preadipocytes in 96-well plates ($1.5{\times}10^4cells/ml$) were cultured with Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS) for 24 h. Then the media were changed to the following 10 different media for 48 h: DMEM containing 10% FBS for the control; the above media containing $5{\mu}M$, $10{\mu}M$ or $15{\mu}M$ of $LaCl_3$, $CeCl_3$ or the mixture of these REE chlorides. The proliferation rate of the cells was measured and compared by a non-isotope method-XTT method. In Experiment 2 the cells in 24-well plates ($1.5{\times}10^4cells/ml$) were cultured in DMEM containing 10% FBS for 7 days until confluent and then were changed to above DMEM containing dexamethasone, methyl-isobutylxanthine and insulin (DMI) for two days. Afterwards the media were changed to the 10 different media with REE supplements as in Experiment 1 and cultured for 6 days. The cells were then harvested for fatty acids analysis by gas chromatography. It was found that supplementation of La (5, 10 and $15{\mu}M$), Ce ($5{\mu}M$ and $15{\mu}M$) and the mixture REE (5, 10 and $15{\mu}M$) stimulated (p<0.05) the proliferation of 3T3-L1 preadipocytes (Experiment 1). In the differentiating 3T3-L1 cells supplementation of La ($5{\mu}M$ and $10{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($5{\mu}M$ and $15{\mu}M$) decreased (p<0.05) the concentration of monounsaturated fatty acids (MUFA) per $10^5cells$, while the supplementation of La ($5{\mu}M$), Ce ($5{\mu}M$) and the mixture REE ($15{\mu}M$) increased (p<0.05) the ratio of saturated fatty acids (SFA) to MUFA. These results indicate that the supplementation of REE to the media may affect proliferation, differentiation and lipogenesis rates of 3T3-L1 cells. However, the effect may depend upon the level or type of REE applied.

• 한국수정란이식학회지
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• 제22권4호
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.