• Toxicological Research
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• 제23권3호
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• pp.271-278
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• 2007

Several recent studies demonstrated the potential of bioengineering using stem cells in regenerative medicine. Adult mesenchymal stem cells (MSCs) have the pluripotency to differentiate into cells of mesodermal origin, i.e., bone, cartilage, adipose, and muscle cells; they, therefore, have many potential clinical applications. On the other hand, stem cells possess a self-renewal capability similar to cancer cells. For safety evaluation of MSCs, in this study, we tested tumorigenecity of canine adipose derived mesenchymal stem cells (cAD-MSCs) using Balb/c-nu mice. In this study, there were no changes in mortality, clinical signs, body weights and biochemical parameters of all animals treated. In addition, there were no significant changes between control and treated groups in autopsy findings. These results indicate that cAD-MSC has no tumorigenic potential under the condition in this study.

• 대한수의학회지
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• 제58권4호
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• pp.201-209
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• 2018

Canine mammary tumors are among the most frequently observed cutaneous tumors in female dogs. Cancer stem cells (CSCs), referred to as tumor-initiating cells, are thought to have properties similar to normal stem cells such as the ability to self-renewal and to differentiate into various cell types. Biological understanding of CSCs and the critical pathways involved in their maintenance are important in research and therapy for mammary tumors. We conducted the present study on sphere formation from REM134 cells by using methylcellulose to produce tumorspheres on a large scale and compared the specific markers of the spheres-formed and plating-cultured REM134 cells. The results revealed that the tumorspheres cultured in methylcellulose had higher seeding density and improved morphology compared to those produced in normal sphere formation medium. Expression levels of stemness markers and CSC-related markers were higher in tumorsphere-forming cells than in plating-cultured cells. Subsequently, we transplanted the tumorsphere-forming and plating-cultured cells into female nude mice to examine their tumorigenic potential. Tumor volume increased rapidly in mice transplanted with tumorsphere-derived cells compared to plating-cultured cells. We observed a novel sphere-forming condition for REM134 cells and showed that REM134 cell tumorspheres can exhibit improved CSC properties.

• Journal of Animal Science and Technology
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• 제65권1호
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• pp.16-31
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• 2023

Cultured meat is a potential sustainable food generated by the in vitro myogenesis of muscle satellite (stem) cells (MSCs). The self-renewal and differentiation properties of MSCs are of primary interest for cultured meat production. MSC proliferation and differentiation are influenced by a variety of growth factors such as insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (FGF-2 and FGF-21), platelet-derived growth factor (PDGF) and hepatocyte growth factor (HGF) and by hormones like insulin, testosterone, glucocorticoids, and thyroid hormones. In this review, we investigated the roles of growth factors and hormones during cultured meat production because these factors provide signals for MSC growth and structural stability. The aim of this article is to provide the important idea about different growth factors such as FGF (enhance the cell proliferation and differentiation), IGF-1 (increase the number of myoblasts), PDGF (myoblast proliferation), TGF-β1 (muscle repair) and hormones such as insulin (cell survival and growth), testosterone (muscle fiber size), dexamethasone (myoblast proliferation and differentiation), and thyroid hormones (amount and diameter of muscle fibers and determine the usual pattern of fiber distributions) as media components during myogenesis for cultured meat production.

• 한국식품영양과학회지
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• 제34권6호
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• pp.805-813
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• 2005

본 연구팀에서는 방사선으로부터 위장관과 면역 조혈계를 보호하기 위하여 당귀, 천궁, 백작약으로부터 새로운 생약복합물 HIM-I을 개발한 바 있다. 본 연구에서는 방사선 방호뿐 아니라 다양한 생체질환의 예방에 있어 면역조혈기능이 중추적인 역할을 한다는 관점에서, HIM-I로부터 면역조혈 기능을 더욱 증진시킨 생약복합조성물 HemoHIM을 제조하여 그 효능을 검증하였다. HIM-I를 에탄을 침지하여 에탄을 분획(HIM-I-E)과 조다당 분획 (HIM-I-P)을 얻은 후, HIM-I 에 조다당 분획을 첨가하여 HemoHIM을 제조하였다. 이렇게 얻은 HemoHIM, HIM-I 및 각 분획에 대하여 재생조직 및 면역계 방호와 회복촉진 효과를 비교 검증하였다. HemoHIM 과 HIM-I는 시험 관내에서 방사선에 의한 DNA 손상을 유의 적으로 억제하고 수산화 라디칼을 소거하는 효과가 있음이 관찰되었으며, HemoHIM과 HIM-I는 거의 비슷한 활성을 나타내었다. 시험 관내 면역 세포 활성화와 골수세포 성장촉 진 실험에서는 HemoHIM이 HIM-I 비하여 높은 활성을 보였으며, 이는 HIM-I에 비해 높은 조다당 함량에 기인한 것으로 보인다. 감마선 조사 마우스를 이용한 생체 보호효과를 살펴본 결과, HemoHIM은 HIM-I와 비슷한 정도의 소장 움 생존율 증가효과를 보였으나, 내재성 비장 조혈세포집락 형성 시험에서는 HemoHIM은 HIM-I보다 높은 효과를 나타내었다. 또한 방사선조사 마우스에서 HemoHTM의 투여는 방사선 조사후 급격히 감소된 말초 혈액내 백혈구 및 림프구수의 회복을 촉진시키고, 생존율을 증대시키는 효과가 관찰되었다. 이상의 결과들은 생약복합물 HIM-I 조다당 분획을 첨가하여 개발한 새로운 생약복합조성물 HemoHIM이 방사선에 의해 유발된 위 장관 및 면역계 조직의 손상을 감소 시켜 생존을 증가시키는 효과가 있음을 제시하였다. 특히 HemoHIM은 HIM-I와 비교하여 재생조직의 산화적 손상억제 효과는 비슷하게 유지되면서도 면역 조혈세포 방호 및 회복촉진 효과가 높은 것으로 관찰되어 방사선 방호제로서 뿐만 아니라 면역조혈기능 증진제로서 유용하게 활용될 수 있을 것으로 사료된다.

• Reproductive and Developmental Biology
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• 제32권1호
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• pp.1-7
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• 2008

본 연구는 돼지 골수에서 존재하는 조혈 줄기 세포 및 전구 세포를 이용해 이종 동물 모델인 태아 마우스 복강 생체 이식을 통하여 돼지 조혈 세포의 이종 조혈 조직에서의 증식과 분화 특성을 규명하였다. 선천성 면역 부전 마우스인 NOD/SCID 마우스 태아 조혈 환경에 돼지 골수 유래 조혈 줄기 세포 및 전구 세포를 이식하고, 이식 후 5주령에 마우스 조혈기관에서의 돼지 조혈 세포의 증식과 분화 특성을 돼지 특이적 항체 면역 염색으로 유세포 분석을 실시한 결과, 마우스 조혈 조직인 골수, 흉선, 간장, 비장 및 림파절에서 돼지 조혈 세포의 분화 및 증식이 관찰되었다. 특히 돼지의 T 면역세포가 골수계 세포에 비해서 높은 chimerism이 관찰되어 태생 초기의 NOD/SCID 조혈 환경에 의한 특이적 T 면역세포의 증식에 적합한 조혈 환경을 제공하고 있다는 사실이 밝혀졌다. 본 마우스 신생 NOSD/SCID 복강 이식 동물 모델을 이용해 돼지 T 면역세포의 분화 발달 연구 및 이종 장기 이식 기전 연구에 좋은 모델로서 활용이 기대된다.

• 대한핵의학회지
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• 제1권1호
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• pp.1-19
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• 1967

The effects of X-ray irradiation and the thyroid gland on the erythropoietic system were studied in the white male rabbits. The total body irradiation was done in doses of 250 r and 500 r to each of 5 rabbits for 10days. The factors were 220KV, 10mA, FLI/4 Cu+1 mmAI(HVL:2.0 mm Cu) 50 cm F.S.D. The thyroid dysfunction was experimentally induced, by giving 2mg of thyroid tablets per kg body weight for 15 days in 5 rabbits for hyperthyroidism and by giving 1.5 mC of $^{131}I$ per kg body weight in another 5 rabbits for hypothyroidism. Fourteen healthy rabbits were used as control. The hematologic changes and ferrokinetic data obtained from $^{59}Fe$ and apparent half survival of the red blood cells obtained from $^{51}Cr$ were compared. Following were the results: A. X-ray irradiated group; 1. There were no significant changes in hematologic findings except for leucopenia. A slight decrease of red blood cells was observed in 500 r irradiated animals. 2. The decreases in the iron turnover rates of the plasma and red blood cells as well as in the red cell renewal rate were found in both groups. A :significant decrease of the red cell iron utilization rate was observed in the 500 r irradiated animals. 3. The apparent half survival times of the red blood cells were slightly, in the 250 r ($12.1{\pm}0.80$ days), and markedly shortened in the 500 r irradiated animals ($9.8{\pm}1.38$ days), the normal being $14.0{\pm}1.6$ days. 4. It appears, therefore, that the anemia caused by X-ray irradiation is due to the inhibition of hemopoietic function and the excess destruction of the red blood cells. B. Thyroid dysfunction group; 1. The slight increases of the red blood cell count and circulating blood volume with the normal serum iron level were observed in the hyperthyroid group, while the decreases of the red and white blood cell counts, hemoglobin and hematocrit values with a marked decrease of the serum iron level in the hypothyroid group. 2. A marked decrease of the plasma iron disappearance rate with increases of plasma iron turnover, red cell iron utilization and red cell iron turnover were observed in the hyperthyroid group, while the marked delay and decreases in the hypothyroid group. 3. The apparent half survival times of the red blood cells were almost the same with the control in the hyperthyroid group, ($14.0{\pm}1.58$ while a marked shortening in the hypothyroid group $10.6{\pm}0.30$. 4. It was reconfirmed that the thyroid hormones bear a close relationship with the erythropoietic system, namely, the latter is stimulated by the former. The lack of the thyroid hormones thus induces the bone marrow depression leading to anemia the major cause of which, therefore, is not hemolysis.

• 대한핵의학회지
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• 제1권2호
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• pp.61-74
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• 1967

In order to confirm whether acquired immunity or resistance can be developed by the repeated hookworm infections, the 150 mature actively moving filariform ancylostoma duodenale larvae obtained from the severe hookworm anemia patients were orally given to 8 healthy volunteers in three divided doses, 50 in each, at 5 day interval. Also the hematological changes as well as several ferrokinetics using $^{59}Fe$ were done and were compared with 10 controls. The clinical symptoms and signs were checked every day for the first 3 weeks and then twice weekly until the end of the experiment. The appearance of the ova in the stool was examined by the formalin ether method and the ova was counted by the Stoll's method. The following laboratory tests were done: 1) Red blood cell count, venous blood hematocrit(micromethod), hemoglobin count (cyanomethemoglobin method) were checked every 5 to 7 day interval. 2) Plasma iron concentration (Barkan's modified method) was determined every 2 to 3 week interval. 3) Radioisotope studies: a) Ferrokinetics: Huff et al and Bothwell's method were applied. Erythropoietic Index (% of normal)=$\frac{Subject's\;turnover/100ml\;whole\;blood{\times}100}{Average\;normal\;turnover/100ml\;whole\;blood}$ of the gastrointestinal absorption of iron: Radioiron($^{59}Fe$) balance b) Quantitative measurement method was applied. c) Determination of the plasma erythropoietin activity: Fried's method was applied. Following were the results: 1) The serum iron level was lower. The red cell volume was decreased, but with relative increase of plasma volume. 2) The plasma iron disappearance time was accelerated and the plasma iron turnover rate was decreased. The red cell iron turnover rate was markedly increased, while all of the red cell iron concentration, circulating red cell iron. plasma iron pool were decreased. The daily iron pool turnover and red cell renewal rate were increased. 3) The erythropoietic index, erythropoietin activity and intestinal absorption of iron($^{59}Fe$) were markedly increased. 4) The infectivity was $9.8{\pm}1.31%$ which was lower than that observed in the single infection. 5) From these observations, it is concluded that the hookworm anemia is essentially iron deficieny in its origin and some immunity acquisition is possible with repeated infections.

• Reproductive and Developmental Biology
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• 제32권1호
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• pp.9-14
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• 2008

조혈 줄기 세포에의 효과적인 유전자 전달은 유전자 치료의 새로운 가능성을 제시할 수 있다. 레트로바이러스를 이용한 유전자 전달 기술은 많은 기초 연구와 임상 시도가 이루어진 대표적인 바이러스이다. 그러나 현재 사용되고 있는 in vitro에서의 조혈 줄기 세포에의 유전자 도입은 조혈 줄기 세포의 분화 유도, 자기 복제 능력과homing 능력의 저하 등 많은 문제점이 있다. 본 연구는 이러한 문제점을 극복하기 위한 방법으로서 마우스의 대퇴골에 직접 레트로바이러스를 이식하는 IBM (Intra-Bone Marrow) 방법을 이용하여 조혈 줄기 세포에의 효과적인 유전자 도입을 시도하였다. IBM 이식 2주 후 마우스의 각 조직을 분석한 결과, 골수뿐 아니라 림파절, 비장, 간장 세포 등에서 유전자가 안정적으로 발현하는 것을 관찰하였다. 또한, $6.4{\pm}2.7%$의 골수조직 존재 조혈줄기/전구세포에서 도입된 유전자가 안정적으로 발현하고 있는 사실을 확인하였다. 본 연구의 결과를 바탕으로 IBM 이식 방법을 이용한 생체 조직 내 레트로바이러스의 유전자 도입은 조혈 줄기 세포를 이용한 유전자 치료에 매우 효과적인 방법이라는 사실을 시사해주고 있다.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.215-220
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• 2007

본 연구는 소 배반포의 내부 세포괴로부터 다능성(pluripotency)을 지닌 배아 줄기 세포(embryonic stem cell) 또는 그 유사 세포를 분리 및 배양함으로써 줄기 세포 관련 분야의 기반 기술을 확립하고자 하였다. 소 체외수정란을 $10{\sim}12$일간 체외배양하여 생산된 부화 배반포를 세포분열이 불활성화된 생쥐 태아 섬유아 세포(mouse embryonic fibroblast, MEF) 위에서 배양하여 콜로니 형성을 유도하였으며, 이들로부터 내부 세포괴 유래의 형태를 지닌 것만을 광학현미경 하에서 물리적으로 분리하여 약 $5{\sim}7$일 간격으로 계대배양을 실시하였다. 이러한 방법을 통하여 배아 줄기 유사 세포의 특성을 40계대 이상 유지하는 2개의 세포주를 확립하였다. 각각의 세포주들은 높은 alkaline phosphatase(AP) 활성을 지니고 있었으며, 형광 면역 염색법과 PCR 기법을 사용하여 Oct-4, Nanog, STAT3, SSEA3 및 SSEA4의 발현을 관찰할 수 있었다. 이러한 결과를 종합하여 볼 때, 본 연구에서는 소 배반포로부터 배아 줄기 세포주를 확립하는 제반 기술이 확립되었다고 판단되며, 향후 관련 분야 연구에 활용될 수 있을 것으로 기대된다.

• BMB Reports
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• 제39권4호
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• pp.426-431
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• 2006

Embryonic stem cells (ESCs), representing a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics, are capable of spontaneous differentiation into cardiomyocytes. The present study sought to define the kinetic characterization of lactate dehydrogenase (LDH) and creatine kinase (CK) of ESC- and neonatal-derived cardiomyocytes. Spontaneously differentiated cardiomyocytes from embryoid bodies (EBs) derived from mouse ESC line (Royan B1) and neonatal cardiomyocytes were dispersed in a buffer solution. Enzymes were extracted by sonication and centrifugation for kinetic evaluation of LDH and CK with spectrophotometric methods. While a comparison between the kinetic properties of the LDH and CK of both groups revealed not only different Michaelis constants and optimum temperatures for LDH but also different Michaelis constants and optimum pH for CK, the pH profile of LDH and optimum temperature of CK were similar. In defining some kinetic properties of cardiac metabolic enzymes of ESC-derived cardiomyocytes, our results are expected to further facilitate the use of ESCs as an experimental model.