• Journal of Microbiology and Biotechnology
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• 제32권2호
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• pp.238-247
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• 2022

Since the skin covers most surfaces of the body, it is susceptible to damage, which can be fatal depending on the degree of injury to the skin because it defends against external attack and protects internal structures. Various types of artificial skin are being studied for transplantation to repair damaged skin, and recently, the production of replaceable skin using three-dimensional (3D) bioprinting technology has also been investigated. In this study, skin tissue was produced using a 3D bioprinter with human skin cell lines and cells extracted from mouse skin, and the printing conditions were optimized. Gelatin was used as a bioink, and fibrinogen and alginate were used for tissue hardening after printing. Printed skin tissue maintained a survival rate of 90% or more when cultured for 14 days. Culture conditions were established using 8 mM calcium chloride treatment and the skin tissue was exposed to air to optimize epidermal cell differentiation. The skin tissue was cultured for 14 days after differentiation induction by this optimized culture method, and immunofluorescent staining was performed using epidermal cell differentiation markers to investigate whether the epidermal cells had differentiated. After differentiation, loricrin, which is normally found in terminally differentiated epidermal cells, was observed in the cells at the tip of the epidermal layer, and cytokeratin 14 was expressed in the lower cells of the epidermis layer. Collectively, this study may provide optimized conditions for bioprinting and keratinization for three-dimensional skin production.

• Journal of Ginseng Research
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• 제35권4호
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• pp.442-448
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• 2011

We previously reported that in vitro anti-platelet activity of tissue-cultured mountain ginseng (TCMG) ethanol extracts show improved efficacy when compared with commercial ginseng products such as Korean red ginseng and Panax ginseng. However, information on the anti-platelet activity of the ethyl acetate fraction from TCMG adventitious roots is limited. Therefore, in this study, we further investigated the effects of an ethyl acetate extract of TCMG (EA-TCMG) adventitious roots on in vitro antiplatelet activity in whole human blood and its effect on peripheral blood flow in mice. We found that EA-TCMG inhibited platelet aggregation with $IC_{50}$ values of 271, 180, and 147 ${\mu}g$/mL induced by collagen, adenosine-5'-diphosphate, and arachidonic acid, respectively. Among the three agonists used, thromboxane $A_2$ formation induced by arachidonic acid was markedly suppressed. Furthermore, EA-TCMG improved the peripheral circulatory disturbance by improving vascular blood flow. In conclusion, these results suggest that ethyl acetate extracts from TCMG adventitious roots might inhibit vascular platelet aggregation and thrombus formation.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1197-1202
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• 2008

Present study investigated the effects of the 70% ethanol extracts of tissue-cultured mountain ginseng (TCMG), Korean red ginseng (KRG), and Panax ginseng (PG) on agonist-induced platelet aggregation and activation in human whole blood. The $IC_{50}$ values for TCMG, KRG, and PG were 1.159, 3.695, and 4.978mg/mL for collagen-induced aggregation, 0.820, 2.030, and 4.743mg/mL for arachidonic acid-induced aggregation, and 1.070, 2.617, and 2.954 mg/mL for ADP-induced aggregation, respectively. Also, this study assessed the effects of the most active extract, TCMG, on markers of platelet activation by determining receptor expression on platelet membranes in healthy subjects, including expression of GPIIb/IIIa-like (PAC-1) and P-selectin (CD62), by flow cytometry. A significant decrease in PAC-l expression (p=0.018) was observed in the presence of TCMG. These results show that TCMG has potent anti-platelet activity.

• 한국동물생명공학회지
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• 제38권1호
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• pp.10-16
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• 2023

Intermuscular fat is essential for enhancing the flavor and texture of cultured meat. Mesenchymal stem cells derived from intermuscular adipose tissues are a source of intermuscular fat. Therefore, as a step towards developing a platform to derive intermuscular fat from mesenchymal stem cells (MSCs) for insertion between myofibrils in cultured beef, an advanced protocol of intermuscular adipose tissue dissociation effective to the isolation of MSCs from intermuscular adipose tissues was developed in cattle. To accomplish this, physical steps were added to the enzymatic dissociation of intermuscular adipose tissues, and the MSCs were established from primary cells dissociated with physical step-free and step-added enzymatic dissociation protocols. The application of a physical step (intensive shaking up) at 5 minutes intervals during enzymatic dissociation resulted in the greatest number of primary cells derived from intermuscular adipose tissues, showed effective formation of colony forming units-fibroblasts (CFU-Fs) from the retrieved primary cells, and generated MSCs with no increase in doubling time. Thus, this protocol will contribute to the stable supply of good quality adipose-derived mesenchymal stem cells (ADMSCs) as a fat source for the production of marbled cultured beef.

• 한국임상수의학회지
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• 제28권5호
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• pp.486-496
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• 2011

조직공학적 인공연골재생에 대한 관심이 증가함에 따라 많은 연구들이 활발히 수행되고 있으나 임상적인 적용의 한계를 극복하기위한 고효능을 보유한 양질의 연골조직생산의 필요성이 증가되고 있다. 인공연골은 자연연골과는 달리 '연골막(perichondrium)'을 포함하고 있지 않기 때문에 장기간 생체 내에 삽입된 후에 서서히 흡수 또는 변형으로 임상적 활용에 한계가 있다고 있다. 이에 본 연구는 양질의 연골조직생산을 목적으로, 세포판 제작기법(cell sheet engineering technique) 을 기반으로 한 인체유래의 배양 연골막(cultured perichondrium)을 이용하여 만든 인공연골막 세포판(cultured perichondrial cell sheet)의 생체 내 특성을 비교 분석하고, 배양된 연골막을 피복하여 고효능화를 유도한 인공연골복합체의 생체내 재생효능 및 조직특성을 비교 평가하고자 하였다. 본 연구에서는 Athymic nude mouse의 피하이식모델(study 1, n = 12)을 이용하여 담체로 hydrogel을 이용한 배양연골막 복합체의 생체내 효능을 분석하였고, 중대형동물의 대량연골 결손시의 재생효능을 평가하기 위하여 개의 무릎연골에 $1{\times}2cm$의 대량연골 결손모델(study 2, n = 12)을 통하여 인공배양세포판을 이식하였다. 이식12주 후 이식편을 회수하여 생화학, 분자생물학 및 면역조직학 분석을 시행한 결과, 배양연골막 복합체의 생체내 효능이 단독이식군에 비해 변형이나 과증식 없이 우수한 결과를 나타내었다. 본 연구의 결과로 토대로 배양연골막을 피복한 인공연골막의 관절내 효과를 규명하여 실제 임상적용을 조기화하는 기반을 제공하고 인공연골의 문제점이었던 변형과 흡수를 줄인 고효능 인공연골 제작기법을 제공하는데 유용할 것으로 기대된다.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.88-94
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• 2005

Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

• 한국식품영양과학회지
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• 제37권8호
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• pp.1018-1024
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• 2008

본 연구에서는 조직배양에 의해 생산된 산삼배양근을 홍삼화하여 부가가치를 향상시키고 새로운 식품소재의 개발을 목적으로 산삼배양근을 압출성형 하였다. 압출성형 산삼 배양근의 일반성분, 조사포닌, 말톨, 산성다당체, 페놀성화합물 등의 화학적 성분과 갈색도, 적색도, 침출특성의 물리적 변화를 살펴보았다. 실험결과 주근의 함량이 높은 홍삼이 총당과 환원당 함량은 산삼배양근과 비교하여 높게 나타났으며, 압출성형 산삼배양근의 총당과 환원당은 증가하였다. 반면 미세근으로 이루어진 산삼배양근의 총아미노산 함량이 주근으로 이루어진 홍삼과 비교하여 높게 측정되었으며, 압출성형 산삼배양근의 아미노산 함량은 다소 감소하는 경향을 나타내었다. 유효성분인 조사포닌 함량은 산삼배양근을 압출성형을 통하여 증가시킬 수 있음을 확인하였으며, 배럴온도 $120^{\circ}C$에서 압출성형한 산삼배양근이 9.60% 가장 높게 측정되었다. 말톨은 홍삼에서만 확인되었으며, 산삼배양근의 경우 압출성형을 통해서도 말톨의 생성은 나타나지 않았다. 산성다당체는 산삼배양근($0.24\;{\mu}g/g$)이 가장 낮았지만 압출성형을 하였을 때 배럴온도 $120^{\circ}C$에서 $0.75\;{\mu}g/g$으로 홍삼의 $0.83\;{\mu}g/g$과 비슷한 수준까지 증가시킬 수 있었다. 페놀성화합물 함량은 압출성형을 통해 감소하였다. 산삼배양근의 갈색도와 적색도는 압출성형으로 증가하였으며, 배럴온도가 증가함에 따라 증가하였다. 갈색도를 기준으로 한 침출특성과 침출속도상수 역시 배럴온도가 증가함에 따라 증가를 확인할 수 있었다. 압출성형은 유효성분인 조사포닌 및 산성다당체 등을 향상시킬 뿐만 아니라 침출력을 향상시켜주므로 향후 고가의 산삼배양근을 더욱 효율적으로 가공할 수 있는 공정으로 확인되었다.

• 한국축산식품학회지
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• 제44권5호
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• pp.1167-1180
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• 2024

This study aimed to investigate effects of chicken age on proliferation and differentiation capacity of muscle satellite cells (MSCs) and to determine total amino acid contents of cultured meat (CM) produced. Chicken MSCs (cMSCs) were isolated from hindlimb muscles of broiler chickens at 5-week-old (5W) and 19-embryonic-day (19ED), respectively. Proliferation abilities (population doubling time and cell counting kit 8) of cMSCs from 19ED were significantly higher than those from 5W (p<0.05). Likewise, both myotube formation area and expression of myosin heavy chain heavy of cMSCs from 19ED were significantly higher than those from 5W (p<0.05). After cMSCs were serially subcultured for long-term cultivation in 2D flasks to produce cultured meat tissue (CMT), total amino acid contents of CMT showed no significant difference between 5W and 19ED chickens (p>0.05). This finding suggests that cMSCs from chicken embryos are more suitable for improving the production efficiency of CM than those derived from young chickens.

• 한국식품조리과학회지
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• 제17권6호
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• pp.549-554
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• 2001

Thermal measurements were made for connective tissues of 5 different fish muscles by using a differential scanning calorimeter(DSC), and connective tissues between muscle fibers and the cross sections of muscle fibers were observed by a light microscope. Red sea bream(cultured and wild) and flounder(cultured, cultured with obosan and wild) were used in this study. It was found that the connective tissues of cultured and frozen fish muscle required less endothermic enthalpy and the endothermic peak temperature was lower than those of wild and fresh ones when they were shrunken and denatured. Therefore, it is likely that the former are more unstable to heat than the latter. The cultured flounder fed with obosan and wild flounder which contained more collagen than cultured flounder and the wild red sea bream showed clear connective tissues between fibers. The cross-section of cultured fish muscle fiber was larger than that of wild one. From these results, collagen content and thermal properties of collagen, cross section of muscle fibers seemed to contribute to the textural difference between wild and cultured fish.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.622-634
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• 1993

Gingiva is remarkly sensitive to certain drugs. Especially, long term use of phentoin, dihydropyrydine (including nifedipine), cyclosporin and other drugs can be lead to pathologic changes in gingival tissue, especially in terms of proliferation of epithelium and connective tissue. Recent study in terms of proliferation of epithelium and connective tissue. Recent study is focused on the inhibition of drug-induced gingival hyperplasia by using medicaments. The purpose of this study was to investigate on the pharmacological effects of nifedipine, retinoic acid and glycyrrhetini acid to the activity in human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and cultured in growth medium added $5{\mu}g/ml$ of nifedipine, $10^{+7}M$ of retinoic acid and glycyrrhetinic acid. The passage number of cultured fibroblasts were between fifth and eighth. The cell morphology was examined by inverted microscope and the cell acitivity was measured by the MTT assay. Nifedipine at the concentration of $5{\mu}g/ml$ was revealed significantly effective to increase the cell activity and lipopolysaccharide was cofactor to increase cell activity in the presence of nifedipine. However, retinoic acid was significantly effective on the globular change of cell morphology and loss of cell process regardless of the presence of nifedipine and LPS. Cell activity was significantly decreased by the glycyrrhetinic acid at the concentration of $10^-M$ regardless of the presence of nifedipine and LPS. These results suggested that the increased cell activity by nifedipine might be modulated by retinoic acid and glycyrrhetinic acid. Further study is needed to clarify on their toxicological effects during cellular modulation and mRNA expression change.