• 대한한방소아과학회지
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• 제14권1호
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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.

• Toxicological Research
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• 제29권4호
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• pp.241-247
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• 2013

This study was carried out to examine the action mechanism of Chamaecyparis obtusa oil (CO) on hair growth in C57BL/6 mice. For alkaline phosphatase (ALP) and ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GT) activities in the skin tissue, at week 4, the 3% minoxidil (MXD) and 3% CO treatment groups showed an ALP activity that was significantly higher by 85% (p < 0.001) and 48% (p < 0.05) and an ${\gamma}$-GT activity that was significantly higher by 294% (p < 0.01) and 254% (p < 0.05) respectively, as compared to the saline (SA) treatment group. For insulin-like growth factor-1 (IGF-1) mRNA expression in the skin tissue, at week 4, the MXD and CO groups showed a significantly higher expression by 204% (p < 0.05) and 426% (p < 0.01) respectively, as compared to the SA group. At week 4, vascular endothelial growth factor (VEGF) expression in the MXD and CO groups showed a significantly higher expression by 74% and 96% (p < 0.05) respectively, however, epidermal growth factor (EGF) expression in the MXD and CO groups showed a significantly lower expression by 66% and 61% (p < 0.05) respectively, as compared to the SA group. Stem cell factor (SCF) expression in the MXD and CO groups was observed by immunohis-tochemistry as significant in a part of the bulge around the hair follicle and in a part of the basal layer of the epidermis. Taking all the results together, on the basis of effects on ALP and ${\gamma}$-GT activity, and the expression of IGF-1, VEGF and SCF, which are related to the promotion of hair growth, it can be concluded that CO induced a proliferation and division of hair follicle cells and maintained the anagen phase. Because EGF expression was decreased significantly, CO could delay the transition to the catagen phase.

• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.587-594
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• 2000

At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

• 생명과학회지
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• 제33권5호
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• pp.445-454
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• 2023

백발화는 자외선, melanin 세포 자극 호르몬(α-MSH), 줄기 세포 인자 성장인자(SCF), Wnt 및 endothelin-1 (ET-1)에 의하여 활성화되는 melanogenesis를 조절하는 신호 전달 경로가 제대로 작동하지 못하여 나타난 결과이다. 백발화를 예방하기 위하여, tyrosinase, tyrosine hydroxylase, tyrosinase-related protein (TRP)-1, TRP-2 및 microphthalmia-associated transcription factor (MITF)에 의하여 조절되는 melanogenesis를 자극하는 효과적인 합성 및 천연 화합물이 있다. 이러한 화합물은 백발화 예방을 위한 잠재성을 지니고 있다. 이 기사는 melanogenesis와 백발화와 관련된 신호 전달 경로에서 최근의 진전뿐 만 아니라 백발화의 문제를 해결하기 위한 핵심적인 전략에 대해 기술한다. 특히, 이글에서는 catalase 및 methionine sulfoxide reductase를 조절하는 항산화제, resveratrol, fisetin, quercetin 및 ginsenoside와 같은 sirtuin (SIRT) 1 activator와 같은 melanin 생성을 촉진하는 잠재적으로 효과적인 치료제에 대하여 설명한다. 또한 estrogen, androgen, progesterone 및 dihydrotestosterone를 포함하는 telomerase 발현 및 activator 뿐만 아니라, corticosteroids, calcineurin restrainer 및 palmitic acid methyl ester와 같은 백반증 억제제에 대하여 논의한다. 더불어 latanoprost, erlotinib, imatinib, tamoxifen, 및 levodopa와 같은 백발화를 억제할 수 있는 화합물에 대해서도 탐구한다. 결론적으로 이 기사는 모발 백발화와 관련된 melanin 생성을 촉진시키는 화합물에 대한 최근의 연구동향을 고찰한다.

• 동의생리병리학회지
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• 제21권1호
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• pp.136-144
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• 2007

Biotae Orientalis has been widely used for treatment of relaxion of smooth muscle, gastrointestinal hemorrhage and alopecia in Oriental Medicine. This experiment examined the effect of an extracts, obtained from the acetone and MeOH extracts of dried or fresh Biotae Orientalis, on hair growing activity of the C57BL/6N mice after topical application to skin. First, We examined on hair growth activity of extracts of Biotae Orientalis compare to control and 1% minoxidil groups. Second, We investigated on the number of hair follicle and mast cells after topical application of extracts of the Biotae Orientalis to skin for 16 day. Third, We investigated immunoreactive density of vascular endothelial growth factor(VEGF), protein kinase C-${\alpha}$(PKC-${\alpha}$) and stem(mast) cell factor(SCF) in skin of C57BL/6N mice by immunohistochemical methods. I fourth investigated changes of subpopulation of splenocytes and thymocytes in C57BL/6N mice for 16day using laser flow cytometry. The results were as follows : Hair growing effect of acetone and MeOH extracts of dried and fresh Biotae Orientalis was observed in 70%, 90% and 60% in hair removed skin area in 16 day respectively. Immunoreactive density of VEGF and PKC-${\alpha}$ in skin of experimental groups was weakly stained compare to control group in 10 day. Immunoreactive density of stem cell factor in skin of experimental group was heavily stained compare to control group in 10 day. Splenic TH/TC Iymphocytes of lived MeOH extracts group significantly increased compare to control group. TH cells in thymic T lymphocytes were increased compare to control group. These experiment suggest that acetone and MeOH extracts of Biotae Orientalis may be used for topical treatment of alopecia areata.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.31-39
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• 2008

손상된 뇌신경조직내에서 신경줄기세포로부터 새로운 신경세포로의 분화가 상당히 제한되어 있어 이것이 손상된 뇌신경조직의 복구가 잘 이루어지지 않는 원인이라 여겨지고 있다. 본 연구에서는 세포배양을 통해 지방조직 중간엽 줄기세포를 도파민성 신경세포와 콜린성 신경세포로 분화를 유도하였다. 중간엽 줄기세포를 신경세포로 분화시키기 위해 N2배양액에 bFGF, EGF, dimethyl sulphoxide (DMSO)와 butylated hydroxyanisole (BHA)를 첨가하여 유도하였다. DMSO와 BHA에 처리된 중간엽 줄기세포가 빠르게 신경세포 모양으로 분화하는 것을 관찰하였으며, 이것은 면역조직학적 염색에서 신경세포 특이 표지인 $\beta$-tubulin III, 별아교세포에 대한 특이 표지인 GFAP, 흰돌기아교세포에 대한 특이 표지인 Gal-C에 대해 양성반응을 나타내었다. RT-PCR 분석에서 배양 단계에 따라 신경세포에 특이적인 표지 인자인 neuro D1, $\beta$-tubulin III, GFAP, nestin 등의 발현을 통해, 중간엽 줄기세포가 신경세포로 분화됨을 확인하였다. 그러나 중간엽줄기세포가 신경세포로 분화된 이후에는 줄기세포 표지인 SCF, C-kit와 stat-3 등은 발현되지 않았다. 또한, 중간엽줄기세포에 bFGF, SHH와 FGF8 등을 처리하면 도파민 신경세포로 분화하였다. 중간엽 줄기세포에 bFGF, RA, Shh를 처리하여 콜린성 신경세포로 분화시켰을 때, 신경세포 특이 표지인 $\beta$-tubulin III와 콜린성 신경 특이 표지인 ChAT에 양성반응를 보였다. 결론적으로 사람 지방조직의 중간엽 줄기세포가 도파민성과 콜린성 신경세포로 분화가 가능하고 이러한 잠재성을 가진 지방 유래 중간엽 줄기세포는 퇴행성 신경질환에 대한 세포 치료제로서 가능성을 제시한다.

• IMMUNE NETWORK
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• 제1권1호
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• pp.87-94
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• 2001

목적 : 제대혈의 조혈모세포 체외확장 시 조혈세포 증폭과 더불어 조혈미세환경의 변화가 일어난다. 이때 제대혈 $CD34^+$ 세포에서 유래되는 지지세포의 계열 분석조혈성장인자 분비능력을 알아보고 지지세포 증식 조건을 확립하여 효과적인 제대혈의 체외증폭을 제시하고자 하였다. 방법 : 제대혈부터 $CD34^+$ 세포를 분리하여 실험에 사용하였다. 무혈청배지에서 각종 조혈성장인자를 다양한 조합으로 첨가하여 배양하였고 증식정도는 현미경으로 관찰하여 배양용기를 점유한 면적 비율로 계산하였다. 세포외간질 단백의 효과를 분석하기 위하여 collagen S, fibronectin, laminin 및 poly-L-ly sine를 미리 coating한 용기에 배양하여 분석하였다. 제대혈 $CD34^+$ 세포를 조혈성장인자의 첨가 없이 3주간 액체배양하였다. 배양 시, 1주, 2주 및 3주에 상층액을 얻어 $-80^{\circ}C$에 보관하였다가 한꺼번에 IL-3, IL-6, GM-CSF, IL-$1{\beta}$ 및 TNF-$\alpha$등을 ELISA 방법으로 내부적으로 분비되는 량을 측정하였다. 분화된 지지세포의 계열을 분석하기 위해 E-selectin, VCAM-1, ICAM-1, PECAM-1, vWF, vimentin 및 CD 14 항체를 이용하여 면역화학염색 후 형광현미경으로 관찰하였다. 결과 : 제대혈 $CD34^+$ 세포 체외증폭시키는 과정에서 배양 4일에 지지세포가 출현하기 시작하여 7-10일이 지나면서 증식하기 시작하였고 14-2 1일 경에 서로 뭉치는 양상을 보여주었다. 제대혈 $CD34^+$ 세포 배양하면서 내부적으로 분비되는 GM-CSF, IL-6의 측정치는 시간이 지남에 따라 증가되었다. 제대혈 $CD34^+$ 세포 체외확장 시 지지세포의 증식 정도는 TPO+FL+SCF+LIF의 조합의 조혈성장인자가 첨가되었을 때 그리고 세포외간질 단백 성분 중 1% poly-L-lysine으로 처리한 경우 가장 효과적이었다. 결론 : 체외 증폭시 제대혈 $CD34^+$ 세포로부터 지지세포가 나타났으며 적절한 조혈성장인자의 첨가나 세포외간질 단백의 첨가에 의해 증폭될 수 있다.

• The Korean Journal of Physiology and Pharmacology
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• 제16권3호
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• pp.167-174
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• 2012

Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-$kit^+$ bone marrow cells (c-$kit^+$ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-$kit^+$ BM cells that give rise to $CD2^+CD8^+$ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-$kit^+$ BM cells than those of controls. And, we compared the ability of the cytotoxicity of $CD2^+CD8^+$ NK cells differentiated by cytokines from c-$kit^+$ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100 : 1 for 4 h once a week. In results, $CD2^+CD8^+$ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-$kit^+$ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-$kit^+$ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.