• 한국가축번식학회지
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• 제17권2호
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• pp.141-148
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• 1993

This studies were carried out to investigate the effects of co-culture with cumulus cells, oviduct epithelial cells and uterine endometrium cells on the in-vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible follicles of diameter 3~5 mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and 10% FCS for 24~48 hrs in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation. The results obtained in these experiments were summarized as follows: 1. The in vitro maturatin and fertilization rate of bovine oocytes co-cultured with cumulus cells in TCM-199 medium were 64.0~74.1% and 40.0~58.6% respectively. And in-vitro fertilization rate of cumulus-enclosed oocytes(55.4%) were significantly(p<0.05) higher than cumulus-denuded oocytes(23.1%). 2. The in-vitro maturatin and fertilization rate of bovine oocytes co-cultured with 1$\times$104 cells/ml, 1$\times$106 cells/ml, 1$\times$108 cells/ml and 1$\times$1015 cells/ml oviduct epithelial cells in TCM-199 medium were 59.3% and 40.7%, 64.0% and 48.0%, 58.3% and 37.5%, 52.0% and 32.0%, respectively. 3. The in-vitro maturation and fertilization rate of bovine oocytes co-cultured with 1$\times$104 cells/ml, 1$\times$106 cells/ml, 1$\times$108 cells/ml and 1$\times$1015 cells/ml uterine endometrium cells in TCM-199 medium were 56.0% and 36.0%, 60.7% and 42.9%, 59.3% and 37.0%, 52.0% and 36.0%, respectively. 4. When the in-vitro fertilized oocytes were co-cultured with cumulus cells, oviduct epithelial cells and uterine endometrium cells, the development rate to be blastocyst was 12.2%, 15.6% and 11.7%, respectively and rates were higher than that of control, 2.1%(P<0.05).

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.127-131
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• 2005

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.119-124
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• 2006

The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

• Clinical and Experimental Reproductive Medicine
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• 제33권4호
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• pp.253-263
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• 2006

목 적: 본 연구에서는 EST Clustering 방법을 이용하여 이전의 연구에서 발굴한 B357 EST의 염기서열을 포함하는 유전자를 동정하고, 이 유전자의 발현을 생쥐의 난소와 정소를 포함한 여러 가지 조직들에서 살펴보고자 하였다. 연구방법: EST Clustering으로 얻어진 전체 염기서열을 5-day-ovary-specific gene-1 (5DOS1)이라고 명명하여 GenBank에 등록하였으며 (AY751521), northern blotting, real-time RT-PCR, in situ hybridization, western blotting, immunohistochemistry 등의 방법을 이용하여 생쥐 난소와 고환의 발달단계에 따라 그 발현양상을 관찰하였다. 결 과: 5DOS1의 전사체는 성장한 정소, 뇌, 근육에서 높게 발현하였으며, 난소의 경우에서는 원시난포시기부터 모든 난자에서 발현하였으며 특히 생후 5일째 높게 발현하였고 그 이후로는 점차 감소하였다. 반면 정소의 경우는 발달과 함께 계속 증가하였으며, 정모세포를 제외한 모든 발달단계별 정자에서 발현함을 관찰하였다. 결 론: 본 연구결과는 5DOS1에 대한 발견과 동정에 대한 첫 보고로써, 유전자 및 단백질이 생쥐의 난소 및 정소의 생식세포에서 발현하는 것을 관찰하였다. 앞으로 생식세포 발생 및 분화에 관련된 5DOS1의 기능에 대한 심층 연구가 더 필요하다고 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제16권2호
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• pp.205-210
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• 1989

To investigate the effects of ovarian cysts on the controlled ovarian hyper-stimulation cycles, 16 patients with 16 paired cycles for IVF-ET were analyzed. These patients had taken both type of cycles, i.e., with cyst(cyst group) and without cyst(control group). Mean diameter of ovarian cysts in cyst group was 18.2mm. There were no significant differences in hormone levels in early follicular phase between two groups. No significant differences were found in total dosage of hMG(IU) administered during the ovarian stimulation $843.8{\pm}123.0$ vs $891.0{\pm}129.8$, serum estradiol level (pg/ml) on the day of hCG administration($1542.8{\pm}1100.6$ vs $1567.5{\pm}1193.0$), the number of aspirated follicles $10.0{\pm}3.4$ vs $11.2{\pm}4.3$ and oocytes $5.3{\pm}3.3$ vs $6.2{\pm}3.1$, the fertilization rate(51.2 % vs 57.2 %) and the cleavage rate(40.5 % vs 52.0 %). Serum estradiol terminal patterns during COH in one group tended to be repeated in the other group. In conclusion, this study suggests that small ovarian cysts do not adversely impact on the controlled ovarian hyperstimulation parameters in IVF - ET program and the presence of small ovarian cyst without concomitant high basal serum estradiol level is not an indication of the cancellation of the controlled ovarian hyperstimulation for IVF-ET.

• Clinical and Experimental Reproductive Medicine
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• 제37권2호
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• pp.153-158
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• 2010

목 적: 과배란유도 후 자궁강내 인공수정 시술시 황체기 보강으로서 경구 미분화 프로게스테론 투여법과 경구 디드로게스테론 투여법의 임상적 효용성을 비교하고자 하였다. 연구방법: 2007년 1월부터 2009년 8월까지 시행된 과배란유도 후 자궁강내 인공수정 시술 183주기를 후향적으로 분석하였다. 과배란유도는 성선자극호르몬 단독 또는 클로미펜과 성선자극호르몬의 병합요법을 사용하였다. 136주기에서는 황체기 보강으로서 경구 미분화 프로게스테론을 하루 300 mg으로 투여하였고 47주기에서는 디드로게스테론을 일일 20 mg으로 투여하였다. 결 과: 여성의 연령, 불임 인자, 성숙난포수 ($\geq$16 mm), 총운동성정자수, triggering 날의 자궁내막 두께는 두 군간 유의한 차이가 없었다. 자궁내 태낭이 확인되는 임상적 임신율은 미분화 프로게스테론 투여군에서 21.3%, 디드로게 스테론 투여군에서 19.1%로 차이가 없었다 (p=0.92). 유산율은 미분화 프로게스테론 투여군에서 다소 높은 경향을 보였으나 통계학적인 차이는 없었다 (34.5% vs. 11.1%, p=0.36). 결 론: 황체기 보강으로서 경구 디드로게스테론 투여법은 경구 미분화 프로게스테론 투여에 비하여 비슷한 임신율과 유산율을 보였다. 그러나 상대적으로 디드로게스테론 투여군의 수가 적어 좀더 많은 환자를 대상으로 한 전향적 연구가 필요하다.

• Journal of Yeungnam Medical Science
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• 제3권1호
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• pp.261-267
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• 1986

치료방사선 분야에서 치료를 제한시키는 요인(dose limiting factor) 중의 하나로 인식되고 있는 피부손상은 그 중요성에 비추어, 임상치료 체계와 같은 방법을 사용한 병리조직학적 연구는 거의 없는 실정이다. 이에 본 연구는 60마리의 백색 마우스를 사용하여 고식적인 분할조사를 통한 급성피부반응의 병리조직학적 변화를 밝히고 후기 합병증의 기선을 추정할 수 있는 기본 자료로 삼고자 하였으며 얻어진 결과는 다음과 같다. 1. 1,000 rad 조사군에서는 성한 표피의 부종만이 관찰되었다. 2. 2,000 rad 조사군에서는 모포와 부속기의 수와 크기가 감소하였고 진피의 부종과 함께 염증 세포의 침윤이 관찰되기 시작하였다. 3. 3,000 rad 조사군에서는 모세혈관 벽의 비후가 관찰되기 시작하였고 부속기의 감소 및 염증 세포의 침윤이 현저히 관찰되었다. 4. 4,000 rad 조사군에서는 더욱 현저한 모세혈관의 비후 및 울혈이 관찰되었다. 5. 5,000 rad 군사군에서는 위의 모든 변화가 증가되었음에도 표피의 완전탈락은 보이지 않았음으로 일반적인 암의 치료양으로는 심한 후기 영향은 고려하지 않아도 좋다고 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권5호
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• pp.708-716
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• 2012

In order to investigate and evaluate the effects of red deer antlers on hair growth in the full-thickness wound healing model, Sprague-Dawley rats were given incision wounds through the full thickness of their dorsal skin and deer antler was applied for 40 days. At specified intervals thereafter (4, 8, 16, 32 and 40 days), the animals were sacrificed and the wound site skins were excised, processed, and sectioned. At post-injury days 16, 32 and 40, longer and more active new hair appeared around the healing wound of antler-treated skin. Histological studies showed that the antler extract markedly increases the depth, size, and number of hair follicles. Expression of IGF-I (insulin-like growth factor) mRNA was detected by RT-PCR and real time RT-PCR. The result showed that the expression of IGF-I (days 16, 32, and 40) was obviously up-regulated in antler-treated skins compared to control skins. Similar results were seen in the ELISA analysis to quantify the IGF-I expression. These results support the notion that wound healing can cause hair growth by enhancing the expression of IGF-I. Deer antler extract appears to have the potential to promote hair growth and could be used in hair growth products.

• 대한약침학회지
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• 제18권4호
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• pp.20-25
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• 2015

Objectives: Recently, thread-embedding therapy (TET) has been widely applied in Korean medicine for cosmetic purposes such as reducing skin wrinkles. An inserted thread was reported to have induced continuous stimulation, followed by support for connective tissue regeneration. However, the potential role of TET in hair-growth has not yet been reported. Methods: We designed this study to evaluate whether TET has a hair-growth-promoting effect. C57 black 6 (C57BL/6) mice were divided into three groups: normal saline-treated, minoxidil-treated, and thread-embedded groups. Normal saline or 5% minoxidil was topically sprayed on the dorsal skin of the mice once a day for 16 days. Medical threads were embedded into the dorsal skin of the mice in a single application. Hair growth activity was evaluated by using dermoscopic and microscopic observations. Sections of the dorsal skin were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), fibroblast growth factor-7 (FGF-7), and fibroblast growth factor-5 (FGF-5) were detected by using immunohistochemical staining. A reverse transcription-polymerase chain reaction (RT-PCR) analysis was adopted to measure the messenger RNA (mRNA) expressions of FGF-7 and FGF-5. Results: TET enhanced anagen development in the hair follicles of C57BL/6 mice. The expressions of BrdU and PCNA, both of which imply active cellular proliferation, were increased by using TET. Moreover, TET increased the expression of FGF-7, an anagen-inducing growth factor, while decreasing the expression of FGF-5, an anagen-cessation growth factor, both at the protein and the mRNA levels. Conclusion: TET enhanced hair re-growth in C57BL/6 mice. TET regulated the expressions of anagen-associated growth factors and activated the proliferation of hair follicular cells in depilated skin lesions. Considering its long-lasting effect, TET may be a good alternative therapeutic for the treatment of alopecia.

• 대한화장품학회지
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• 제40권4호
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• pp.413-421
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• 2014

코르티코트로핀분비인자(Corticotropin-releasing factor)는 스트레스 반응에 관여하는 호르몬으로, 최근 스트레스가 탈모와 같은 피부질환에 영향을 미친다는 보고들이 많아지고 있다. 보고에 따르면, 사람 모낭 배양에서 코르티코트로핀분비인자는 길이생장을 억제하며, 모낭의 조기퇴행을 유도하고 모기질각질형성세포(hair matrix keratinocyte)의 세포사멸을 촉진시킨다. 본 연구에서는 코르티코트로핀분비인자가 모발성장과 모주기조절에 핵심적으로 역할하는 모유두세포에 미치는 영향에 대해 알아보고자 했다. 시상하부-뇌하수체-부신축의 주요 스트레스호르몬들인 코르티코트로핀분비인자, 부신피질자극호르몬, 그리고 코르티솔을 사람 모유두세포에 처리하였다. 흥미롭게도, 코르티코트로핀분비인자가 모발성장과 관련된 사이토카인(KGF, Wnt5a, $TGF{\beta}-2$, Nexin)의 발현을 변화시키는 것을 관찰하였으며, 세포 내 cAMP의 수준을 증가시켰고, 수용체의 발현을 억제시켰다. 이러한 변화는 수용체의 길항제인 antalarmin과 astressin2B, 또는 PKA 억제제의 전처리로 인해 막을 수 있었다. 코르티코트로핀분비인자는 cAMP/PKA경로를 통해 POMC의 발현을 유도하는데, 사람 모유두세포에서도 이 호르몬의 처리가 POMC mRNA의 발현을 증가시키는 것을 확인할 수 있었으나 부신피질자극호르몬의 변화는 western blot으로는 확인할 수 없었다. 이러한 결과들을 바탕으로, 코르티코트로핀분비인자가 그 수용체를 통해 사람 모유두세포 내 모발성장 관련 사이토카인의 발현을 조절함을 확인하였으며, 이는 코르티코트로핀분비인자의 수용체 길항제가 스트레스성 탈모환자를 위한 치료제 혹은 화장품 소재로써 활용될 수 있음을 보여준다.