• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.327-333
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• 2006

Future cell-based therapies such as tissue engineering will benefit from a source of autogenous pluripotent stem cells. There are embryonic stem cells (ESC) and autologous adult stem cells, two general types of stem cells potentilally useful for these applications. But practical use of ESC is limited due to potential problems of cell regulation and ethical considerations. To get bone marrow stem cells is relatively burden to patients because of pain, anesthesia requirement. The ideal stem cells are required of such as the following advantages: easy to obtain, minimal patient discomfort and a capability of yielding enough cell numbers. Adipose autologus tissue taken from intraoral fatty pad or abdomen may represent such a source. Our study designed to demonstrate the ability of human adipose tissue-derived stromal cells (hATSC) from human abdominal adipose tissue diffentiating into osteocyte and adipocyte under culture in vitro conditions. As a result of experiment, we identified stromal cell derived adipose tissue has the multilineage potentiality under appropriate culture conditions. And the adipose stromal cells expressed several mesenchymal stem cell related antigen (CD29, CD44) reactions. Secondary, we compared the culture results of a group of hATSC stimulated with TGF-${\beta}$1, bFGF with a hATSC group without growth factors to confirm whether cytokines have a important role of the proliferation in osteogenic differentiation. The role of cytokines such as TGF-${\beta}$1, bFGF increased hATSC's osteogenic differentiation especially when TGF-${\beta}$1 and bFGF were used together. These results suggest that adipose stromal cells with growth factors could be efficiently available for cell-based bone regeneration.

• 대한치과교정학회지
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• 제27권5호
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• pp.683-696
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• 1997

성체의 상처 치유과정중 반흔조직 형성에 수종의 성장인자가 관련이 있는 것으로 보고되고 있으나, 태자의 피부상처가 반흔형성없이 치유되는 기전에 관한 성장인자의 역할은 아직 알려져 있지 않다. 따라서 본 연구의 목적은 토끼태자의 상처 치유과정에서 반흔조직의 형성과 관련이 있는 수종 성장인자의 분포를 조사하기 위한 것이다. 뉴질랜드산 흰 토끼를 임신 3기의 중반인 24일째에 자궁절개술을 시행하고 태자에 인위적으로 구순열상을 만든 후, 봉합한 군(봉합군)과 봉합하지 않은 군(비봉합군), 정상대조군(sham operated control group)으로 나누고 이들을 각각 수술후 1, 2, 3, 5, 7일째에 희생시켜 상처치유에 대한 육안관찰 소견과 H & E 염색 소견 및 $TGF-{\beta}1,\;TGF-{\beta}2$, PDGF, bFGF의 면역조직화학적 염색 소견을 관찰하여 다음과 같은 결론을 얻었다. 1. 봉합군과 비봉합군에서 전기간동안 염증반응과 반흔조직의 형성 그리고 신생 혈관의 형성 증가는 없었다. 2. 비봉합군의 재상피화가 봉합군에 비해 다소 느렸다. 3. bFGF는 정상대조군, 봉합군과 비봉합군에서 차이가 없었다. 4. PDGF는 봉합군과 비봉합군에서 1, 2일군에서 증가하였다가 그후 감소하여 정상대조군과 차이가 없었다. 5. $TGF-{\beta}$는 봉합군과 비봉합군에서 1, 2일군에서 약간 증가하였다가 그후 감소하여 정상대조군과 차이가 없었다. $TGF-{\beta}1$에 비하여 $TGF-{\beta}2$의 검출양이 많았다.

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.656-662
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• 2001

Angiogenesis is known as a crucial process in the growth and spreading of tumor cells. Accordingly, the effective inhibition of this process would appear to be a promising way to cure angiogenesis-related diseases, including cancer. This study demonstrates that acalycixenolide E (AX-E) from the marine organism Acalycigorgia inermis exhibits a potent anti-angiogenic activity both in vitro and in vivo. AX-E inhibits the bFGF-induced proliferation of HUVECs in a dose dependent manner, along with the bFGF-induced migration, invasion, and tube formation of HUVECs. Moreover, AX-E potently inhibits the in vivo neovascularization of the chorioallantoic membranes (CAMs) of growing chick embryos. interestingly, AX-E suppresses the expression of metalloproteases 2 and 9, yet shows no effect on their activities. The novel chemical structure and potent anti-angiogenic activity of AX-E will be of great value in elucidating the molecular mechanism of angiogenesis as well as in the development of a novel anti-angiogenic drug.

• 대한수의학회지
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• 제52권2호
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• pp.99-104
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• 2012

The present study was performed to evaluate the relationship between the neurotoxicity of acrylamide and the differential gene expression pattern in mice. Both locomotor test and rota-rod test showed that the group treated with higher than 30 mg/kg/day of acrylamide caused impaired motor activity in mice. Based on cDNA microarray analysis of mouse brain, myelin basic protein gene, kinesin family member 5B gene, and fibroblast growth factor (FGF) 1 and its receptor genes were down-regulated by acrylamide. The genes are known to be essential for neurofilament synthesis, axonal transport, and neuroprotection, respectively. Interestingly, both FGF 1 and its receptor genes were down-regulated. Genes involved in nucleic acid binding such as AU RNA binding protein/enoyl-coA hydratase, translation initiation factor (TIF) 2 alpha kinase 4, activating transcription factor 2, and U2AF 1 related sequence 1 genes were down-regulated. More interesting finding was that genes of both catalytic and regulatory subunit of protein phosphatases which are important for signal transduction pathways were down-regulated. Here, we propose that acrylamide induces neurotoxicity by regulation of genes associated with neurofilament synthesis, axonal transport, neuro-protection, and signal transduction pathways.

• Reproductive and Developmental Biology
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• 제32권4호
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• pp.263-267
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• 2008

This study was conducted to compare the expression pattern of the specific factors associated with pregnancy and angiogenesis during early pregnancy in Hanwoo. Synchronized female Hanwoo ($4{\sim}6$ year-old) were inseminated artificially. After 10 weeks after artificial insemination (AI), the pregnancy was tested by rectal palpation method. Three pregnant and non-pregnant Hanwoo were used in this experiment, respectively. The plasma progesterone level was measured by ELISA. Western blot analysis was performed to detect the expression of pregnancy associated glycoprotein (PAG) or angiogenic factors (VEGF, B-FGF, ANP-1, and TIE-2). The plasma P4 level was increase gradually in pregnant group and maintained high level. The concentration of PAG was significantly higher from $5^{th}$ weeks in pregnant group compared to that of non-pregnant group (p<0.05). The concentrations of the VEGF (p<0.05), B-FGF (p<0.05), and ANP-1 (p<0.05) were significantly increased from $6^{th}\;or\;7^{th}$ week after AI in pregnant group, respectively. And the intensity of TIE-2, ANP-1 receptor, was well matched with ANP-1 (p<0.05). Taken together, it can be postulated that the blood vessels connected with fetus and dam were formed dramatically around 40 days after AI, because the expression levels of the angiogenic factors were increased significantly from this time in pregnant Hanwoo.

• 한국발생생물학회지:발생과생식
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• 제11권3호
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• pp.167-177
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• 2007

본 연구에서는 사람의 지방조직(human adipose tissue-derived stem cells, HAD), 탯줄(human umbilical cordderived stem cells, HUC), 그리고 양막(human amnion-derived stem cells, HAM)유래 줄기세포를 분리하여 세포의 형태 및 성장속도를 비교하고, 역전사 중합효소 연쇄반응과 면역세포화학 염색법을 이용하여 유전자와 단백질 발현을 비교 분석하였다. 또한 지방유래 줄기세포를 이용하여 심장근육세포로의 분화를 유도하였다. 본 연구 결과, 탯줄과 양막유래 줄기세포의 형태는 매우 유사하였으며, 지방유래 줄기세포의 형태는 다른 것으로 나타났다. 분열시간은 탯줄유래 줄기세포가 가장 빨랐으나 총 분열 횟수는 양막유래 줄기세포와 같았으며, 지방유래 줄기세포의 총 분열횟수가 가장 많았다. 세 종류 줄기세포의 유전자와 단백질 발현은 비슷한 양상을 나타냈다. 지방세포, 골세포, 연골세포로의 분화 유도 결과 세 종류의 줄기세포 모두 분화 유도되었다. 또한, 심장세포 특이 유전자의 발현 분석 결과 세 종류의 줄기세포에서 유사한 발현 양상을 나타냈다. 이 중 지방유래 줄기세포를 24시간 동안 $10\;{\mu}M$ 5-azacytidine 처리 후 기본 배양액에서 4주 동안 배양하거나 또는 5-azacytidine 처리 후 bone morphogenic protein-2(BMP-2)와 fibroblast growth factor-10(FGF-10) 또는 BMP-4와 FGF-4 또는 BMP-4와 FGF-8이 첨가된 배양액으로 4주 동안 배양하여 심근세포로의 분화를 유도하였다. 분화 유도 후 심장세포 특이 유전자 발현을 분석 결과 cardiac myosin light chain-1(Cmlc-1)과 L-type calcium channel ${\alpha}1C$ subunit(${\alpha}1C$) 유전자의 발현이 증가하였다. 그러나 troponin T(TnT), troponin I(TnI) 그리고 potassium channel Kv4.3 subunit (Kv4.3) 유전자의 발현은 증가하지 않았다. 본 연구 결과, 지방, 탯줄 및 양막유래 줄기세포는 특성이 매우 유사한 것으로 나타났으며, 심장 질환 치료를 목적으로 하는 세포 치료에 이용될 수 있을 것으로 사료된다. 또한, 적절한 배양조건 하에서 성장인자와 cytokine들을 처리하여 심장세포로의 분화 유도가 이루어진다면 임상적용에 유용한 세포로 사용될 수 있을 것으로 사료된다.

• 한국발생생물학회지:발생과생식
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• 제12권2호
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• pp.159-168
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• 2008

Nanog is a newly identified member of the homeobox family of DNA binding transcription factors that functions to maintain the undifferentiated state of stem cells. However, molecular mechanisms underlying the function of Nanog remain largely unknown. To elucidate the regulatory roles of Nanog involved in maintenance of P19 embryonal carcinoma (EC) stem cells, we transfected three small interfering RNA (siRNA) duplexes targeted against different regions of the Nanog gene into P19 cells. The Nanog siRNA-100 duplexes effectively decreased the expression of Nanog up to 30.7% compared to other two Nanog siRNAs, the Nanog siRNA-400 (67.9 %) and -793 (53.0%). When examined by RT-PCR and real-time PCR, the expression of markers for pluripotency such as Fgf4, Oct3/4, Rex1, Sox1 and Yes was downregulated at 48 h after transfection with Nanog siRNA-100. Furthermore, expression of the ectodermal markers, Fgf5 and Isl1 was reduced by Nanog knockdown. By contrast, the expression of other markers for pluripotency such as Cripto, Sox2 and Zfp57 was not affected by Nanog knockdown at this time. On the other hand, the expression of Lif/Stat3 pathway molecules and of the endoderm markers including Dab2, Gata4, Gata6 and the germ cell nuclear factor was not changed by Nanog knockdown. The results of this study demonstrated that the knockdown of Nanog expression by RNA interference in P19 cells was sufficient to modulate the expression of pluripotent markers involved in the self-renewal of EC stem cells. These results provide the valuable information on potential downstream targets of Nanog and add to our understanding of the function of Nanog in P19 EC stem cells.

• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.267-271
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• 1997

We developed a dermal equivalent (DE) which was engineered using human dermal fibroblasts and a matrix of collagen gel. The in vitro construction of the DE was accomplished by casting a porcine collagen type I solution plus concentrated medium with isolated and cultured fibroblasts. These constructs were attached to culture dishes or left floating in culture medium. Contraction of attached gels results in decreased gel thickness without a change in gel diameter, and contraction of floating gels results in decreased gel thickness and diameter. After contraction, there was no increase in cell number in floating gels, but cells in attached gels began to increase after about 4 days of the lag phase in cell growth curve. At this lag phase, addition of fibroblast growth factor (FGF) at a concentration of $0.1{\mu}$/ml promoted cell proliferation in the attached collagen gels, but no effect in floating gels. These results indicate that the method of contraction had an influence on the extracellular matrix (ECM) organization, and this influenced not only cell growth but also fibroblast responsiveness to FGF. This suggests that attached collagen gel is more suitable as a dermal equivalent than the floating gel. And the final contracted area of attached gel is much larger than that of the floating gel since floating gel is contracted in all directions but attached gel is contracted only vertically.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.80-80
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• 2003

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms, accompanied by marked cell death in the striatum and cortex. Stereotaxic injection of quinolinic acid (QA) into striatum results in a degeneration of GABAergic neurons and exhibits abnormal motor behaviors typical of the illness. The objective of this study was carried out to obtain basic information about whether parthenogenetic mouse embryonic stem (PmES) cells are suitable for cell replacement therapy of HD. To establish PmES cell lines, hybrid F1 (C57BL/6xCBA/N) mouse oocytes were treated with 7% ethanol for 5 min and cytochalasin-B for 4 hr to initiate spontaneous cleavage. Thus established PmES cells were induced to differentiate using bFGF (20ng/ml) followed by selection of neuronal precursor cells for 8 days in N2 medium. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days, then a final differentiation step in N2 medium for 7 days. To establish recipient animal models of HD, young adult mice (7 weeks age ICR mice) were lesioned unilaterally with a stereotaxic injection of QA (60 nM) into the striatum and the rotational behavior of the animals was tested using apomorphine (0.1mg/kg, IP) 7 days after the induction of lesion. Animals rotating more than 120 turns per hour were selected and the differentiated PmES cells (1$\times$10$^4$cells/ul) were implanted into striatum. Four weeks after the graft, immunohistochemical studies revealed the presence of cells reactive to anti-NeuN antibody. However, only a slight improvement of motor behavior was observed. By Nissl staining, cell mass resembling tumor was found at the graft site and near cortex which may explain the slight behavioral improvement. Detailed experiment on cell viability, differentiation and migration explanted in vivo is currently being studied.

• Applied Microscopy
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• 제37권2호
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• pp.93-102
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• 2007

분비백혈구단백분해효소억제제 (SLPI)와 여러 성장인자들은 상처 감염이나 박테리아 침입 시 일어나는 염증반응에 서로 관계가 있지만, 대식세포에서 LPS 자극시 SLPI와 VEGF, bFGF, PDGF 등과 같은 성장 인자들의 발현관계에 대해서는 아직까지 알려진 연구 결과가 없다. 따라서 본 연구는 대식세포 세포주로 알려진 RAW264.7 세포에 SLPI 발현의 적정농도인 LPS에 반응하는 SLPI 및 성장 인자들과 세포외부기질이 발현을 규명하고자 하였다. 역전사효소 중합반응(RT-PCR)과 면역학적 단백질 검출법(Western blotting)은 LPS 처리 후 SLPI와 몇몇 성장인자들 (VEGF, bFGF, PDGF)와 제1형 아교질 mRNA와 SLPI 단백질의 검출을 위해 수행하였다. RAW264.7 세포 주를 mL 당 100 ng의 LPS에 각각 30분, 60분, 90분, 24시간, 48시간동안 노출시켰다. RT-PCR 결과 SLPI mRNA는 시간이 지남에 따라 점점 발현 양이 증가하였고 VEGF와 PDGF mRNA는 초기에 높은 발현 양상을 보였다. 그러나 bFGF와 I형 아교질의 발현은 매우 미약하게 나타났다. SLPI 단백질 발현 역시 mRNA 수준과 마찬가지로 증가하는 양상을 보였는데, 배양액내의 SLPI 단백질양은 전체적으로 감소하는 경향을 보였다. 또한 광학현미경 관찰과 주사전자현미경 관찰결과, LPS가 RAW264.7 세포주의 형태학적인 변화를 일으킴을 확인하였다. 본 결과를 종합하면 SLPI 발현증가의 적정 농도라 생각되는 100ng의 LPS에 의해서 발현되는 VEGF나 PDGF는 SLPI의 발현에 관계가 있는 것으로 생각되지만 추후에 이들 인자들의 단백질이나 유전자 도입을 통하여 발현 관계를 명확히 확인해야 하는 추가실험이 진행되어야 할 것이다.하게 된다.토끼 면역항체를 선모충유충 조직항원에 반응시켰을 때 충체의 표피와 기저층 그리고 EIM 및 stichocyte의 ${\alpha}_0\;{\alpha}_1$ 과립에 황금입자가 표지되었다. 따라서 1일 동안 배설되는 분비배설항원은 선모충 유충의 표피와 stichocyte의 ${\alpha}_0\;{\alpha}_1$ 과립에서 유도되는 반면에 3일 동안 배설되는 분비배설항원은 표피와 stichocyte의 ${\alpha}_0$ 과립에서 유도되고, 선모충유충 감염후 1주, 4주에 실험쥐에서 형성되는 감염항체는 선모충의 표피와 기저층 그리고 EIM에서 분비되는 항원에 의하여 생성된다. 이상의 결과로 선모충의 분비배설항원과 감염항원은 선모충 유충의 표피와 EIM및 stichocyte의 ${\alpha}_0\;{\alpha}_1$ 과립에서 유도되며 이들은 45 kDa 단백을 포함하고 있는 것으로 생각된다.성하고 있는 세포들에는 세포질이 어두운 세포와 밝은 세포가 있었으며, 세포질내에는 전자밀도가 높은 분비과립이 관찰되었다. 전체적인 특징은 눈물샘분비세포 중 장액세포의 것과 비슷하였으나, 과립의 크기는 작았다. 분비관을 구성하는 세포들 사이에도 연접복합체가 매우 잘 발달되어 있었다. 샘포에서 사이관으로 이행되는 곳에서도 샘포세포와 사이관세포 사이에서도 연접복합체가 관찰되었다. 분비관세포의 분비과립 가운데는 중심부분에 전자밀도가 더 높은 중심을 가진 다른 모양의 과립이 관찰되기도 하였다. 의해 사망한 환자는 없었다. 결 론: 자궁경부암 환자에 항암화학요법과 동시에 외부 방사선조사와 고선량률의 강내조사를 시행한 결과 독성이 심하지 않고 국소제어율과 단기 생존율이 양호하여 안전하고 효율적인 치료방법으로 생각된다.