• 한국발생생물학회지:발생과생식
• /
• 제16권4호
• /
• pp.339-351
• /
• 2012

Fetal bovine serum (FBS) is the most frequently used serum for the cultivation of mammalian cells. However, since animal-derived materials might not be appropriate due to safety issues, allogeneic human serum (HS) has been used to replace FBS, particularly for the culture of human cells. While there has been a debate about the advantages of HS, its precise effect on human adult stem cells have not been clarified. The present study aimed to investigate the effect of HS on the human eyelid adipose stem cells (HEACs) in vitro. When HEACs were cultivated in a medium containing 10% HS, many cells moved into several spots and aggregated there. The phenomenon was observed as early as 9 days following 10% HS treatment, and 12 days following 5% HS plus 5% FBS treatment. However, the aggregation was never observed when the same cells were cultivated with 10% FBS or bovine serum albumin. To examine whether cell density might affect the aggregation, cells were seeded with different densities on 12-well dish. Until the beginning of aggregation, cells seeded at low densities exhibited the longest culture period of 16 days whereas cells seeded at high densities showed the shortest period of 9 days to form aggregation. The number of cells was $15.1{\pm}0.2{\times}10^4$ as the least for the low density group, and $29.3{\pm}2.8{\times}10^4$ as the greatest for the high density group. When human cord blood serum or normal bovine serum was examined for the same effect on HEACs, interestingly, cord blood serum induced the aggregation of cells whereas bovine serum treatment has never induced. When cells were cultivated with 10% HS for 9 days, they were obtained and analyzed by RT-PCR. Compared to FBS-cultivated HEACs, HS-cultivated HEACs did not express VIM, and less expressed GATA4, PALLD. On the other hand, HS-cultivated HEACs expressed MAP2 more than FBS-cultivated HEACs. In conclusion, human adult stem cells could move and form aggregates by the treatment with human body fluids.

• The Plant Pathology Journal
• /
• 제40권1호
• /
• pp.48-58
• /
• 2024

The oldest and most extensively cultivated form of millet, known as pearl millet (Pennisetum glaucum (L.) R. Br. Syn. Pennisetum americanum (L.) Leeke), is raised over 312.00 lakh hectares in Asian and African countries. India is regarded as the significant hotspot for pearl millet diversity. In the Indian state of Haryana, where pearl millet is grown, a new and catastrophic bacterial disease known as stem rot of pearl millet spurred by the bacterium Klebsiella aerogenes (formerly Enterobacter) was first observed during fall 2018. The disease appears in form of small to long streaks on leaves, lesions on stem, and slimy rot appearance of stem. The associated bacterium showed close resemblance to Klebsiella aerogenes that was confirmed by a molecular evaluation based on 16S rDNA and gyrA gene nucleotide sequences. The isolates were also identified to be Klebsiella aerogenes based on biochemical assays, where Klebsiella isolates differed in D-trehalose and succinate alkalisation tests. During fall 2021-2023, the disease has spread all the pearl millet-growing districts of the state, extending up to 70% disease incidence in the affected fields. The disease is causing considering grain as well as fodder losses. The proposed scale, consisting of six levels (0-5), is developed where scores 0, 1, 2, 3, 4, and 5 have been categorized as highly resistant, resistant, moderately resistant, moderately susceptible, susceptible, and highly susceptible disease reaction, respectively. The disease cycle, survival of pathogen, and possible losses have also been studied to understand other features of the disease.

• 한국산림과학회지
• /
• 제99권1호
• /
• pp.57-61
• /
• 2010

본 연구는 난대산림연구소의 제주시험림에 있는 붉가시나무(Quercus acuta)에 대한 개체목의 수간곡선식 추정 및 간재적을 추정하기 위하여 수행되었다. 최적의 추정식을 선택하기 위하여 Max and Burkhart식, Kozak식 및 Lee식을 적용하여 각 식의 직경 추정에 대한 검정 통계량 및 실측치와 추정치간의 오차분포를 검증하였다. 그 결과 Max and Burkhart식 및 Lee식이 특정 구간에서 과대치 또는 과소 추정치를 보인데 반하여 Kozak식은 전구간에서 고른 분포를 보였다. 추정력이 가장 좋은 Kozak식을 활용하여 수피포함 재적표를 작성하였다.

• 대한음성언어의학회:학술대회논문집
• /
• 대한음성언어의학회 2003년도 제19회 학술대회
• /
• pp.183-183
• /
• 2003

Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.

• Reproductive and Developmental Biology
• /
• 제36권1호
• /
• pp.21-26
• /
• 2012

Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

• BMB Reports
• /
• 제54권10호
• /
• pp.505-515
• /
• 2021

Human pluripotent stem cells (hPSCs) include human embryonic stem cells (hESCs) derived from blastocysts and human induced pluripotent stem cells (hiPSCs) generated from somatic cell reprogramming. Due to their self-renewal ability and pluripotent differentiation potential, hPSCs serve as an excellent experimental platform for human development, disease modeling, drug screening, and cell therapy. Traditionally, hPSCs were considered to form a homogenous population. However, recent advances in single cell technologies revealed a high degree of variability between individual cells within a hPSC population. Different types of heterogeneity can arise by genetic and epigenetic abnormalities associated with long-term in vitro culture and somatic cell reprogramming. These variations initially appear in a rare population of cells. However, some cancer-related variations can confer growth advantages to the affected cells and alter cellular phenotypes, which raises significant concerns in hPSC applications. In contrast, other types of heterogeneity are related to intrinsic features of hPSCs such as asynchronous cell cycle and spatial asymmetry in cell adhesion. A growing body of evidence suggests that hPSCs exploit the intrinsic heterogeneity to produce multiple lineages during differentiation. This idea offers a new concept of pluripotency with single cell heterogeneity as an integral element. Collectively, single cell heterogeneity is Janus-faced in hPSC function and application. Harmful heterogeneity has to be minimized by improving culture conditions and screening methods. However, other heterogeneity that is integral for pluripotency can be utilized to control hPSC proliferation and differentiation.

• 생체재료학회지
• /
• 제22권4호
• /
• pp.271-278
• /
• 2018

Background: The chondrogenic differentiation of mesenchymal stem cells (MSCs) is regulated by many factors, including oxygen tensions, growth factors, and cytokines. Evidences have suggested that low oxygen tension seems to be an important regulatory factor in the proliferation and chondrogenic differentiation in various MSCs. Recent studies report that synovium-derived mesenchymal stem cells (SDSCs) are a potential source of stem cells for the repair of articular cartilage defects. But, the effect of low oxygen tension on the proliferation and chondrogenic differentiation in SDSCs has not characterized. In this study, we investigated the effects of hypoxia on proliferation and chondrogenesis in SDSCs. Method: SDSCs were isolated from patients with osteoarthritis at total knee replacement. To determine the effect of oxygen tension on proliferation and colony-forming characteristics of SDSCs, A colony-forming unit (CFU) assay and cell counting-based proliferation assay were performed under normoxic (21% oxygen) or hypoxic (5% oxygen). For in vitro chondrogenic differentiation, SDSCs were concentrated to form pellets and subjected to conditions appropriate for chondrogenic differentiation under normoxia and hypoxia, followed by the analysis for the expression of genes and proteins of chondrogenesis. qRT-PCR, histological assay, and glycosoaminoglycan assays were determined to assess chondrogenesis. Results: Low oxygen condition significantly increased proliferation and colony-forming characteristics of SDSCs compared to that of SDSCs under normoxic culture. Similar pellet size and weight were found for chondrogensis period under hypoxia and normoxia condition. The mRNA expression of types II collagen, aggrecan, and the transcription factor SOX9 was increased under hypoxia condition. Histological sections stained with Safranin-O demonstrated that hypoxic conditions had increased proteoglycan synthesis. Immunohistochemistry for types II collagen demonstrated that hypoxic culture of SDSCs increased type II collagen expression. In addition, GAG deposition was significantly higher in hypoxia compared with normoxia at 21 days of differentiation. Conclusion: These findings show that hypoxia condition has an important role in regulating the synthesis ECM matrix by SDSCs as they undergo chondrogenesis. This has important implications for cartilage tissue engineering applications of SDSCs.

• Journal of Periodontal and Implant Science
• /
• 제53권6호
• /
• pp.467-477
• /
• 2023

Purpose: The purpose of this study was to evaluate the regenerative capacity of stem cells combined with bone graft material and a collagen matrix in rabbit calvarial defect models according to the type and form of the scaffolds, which included type I collagen matrix and synthetic bone. Methods: Mesenchymal stem cells (MSCs) were obtained from the periosteum of participants. Four symmetrical 6-mm-diameter circular defects were made in New Zealand white rabbits using a trephine drill. The defects were grafted with (1) group 1: synthetic bone (β-tricalcium phosphate/hydroxyapatite [β-TCP/HA]) and 1×10⁵ MSCs; (2) group 2: collagen matrix and 1×10⁵ MSCs; (3) group 3: β-TCP/HA, collagen matrix covering β-TCP/HA, and 1×10⁵ MSCs; or (4) group 4: β-TCP/HA, chipped collagen matrix mixed with β-TCP/HA, and 1×10⁵ MSCs. Cellular viability and cell migration rates were analyzed. Results: Uneventful healing was achieved in all areas where the defects were made at 4 weeks, and no signs of infection were identified during the healing period or at the time of retrieval. New bone formation was more evident in groups 3 and 4 than in the other groups. A densitometric analysis of the calvarium at 8 weeks post-surgery showed the highest values in group 3. Conclusions: This study showed that the highest regeneration was found when the stem cells were applied to synthetic bone along with a collagen matrix.

• 정신신체의학
• /
• 제10권1호
• /
• pp.8-17
• /
• 2002

연구목적: 본 조혈모세포 이식술의 광범위한 사용 및 시술 후 생존율의 증가와 함께 조혈모세포 이식환자의 심리적 적응에 대한 관심이 증가하였다. 이에 본 연구에서는 조혈모세포 이식환자의 스트레스 대처방식과 발현될 수 있는 정신과적 증상에 대해 알아보고자 하였다. 방법: 동종 조혈모세포 이식을 전후하여 환자들의 심리적 스트레스 증상을 평가하였으며 스트레스 대처방식에 심리적 변인의 차이에 대해 알아보았다. 29명의 조혈모세포 이식 환자가 본 연구에 참여하여 조혈모세포 이식 직전과 직후에 평가를 시행하였으며, 30명의 정상 대조군이 본 연구에 참여하였다. 평가방법으로는, 면담을 통한 정신상태검사와 여러 가지 표준화된 설문지(스트레스 대처방식 척도, 스트레스 지각 척도, Short-Form 36 Health Survey)를 사용하였다. 결과: 환자군에서 정상 대조군에 비해 우울(p<0.001) 이나 불안(p=0.011) 의 정도가 유의하게 높았으나, 환자군의 조혈모세포 이식 전후에 우울과 불안은 차이가 없었다 또한 소극적인 대처방식을 사용하는 환자군에서 우울(p=0.046) 이나 불안(p<0.001)의 정도가 유의하게 높았다. 결론: 본 연구를 통해 조혈모세포 이삭 환자들이 정상 대조군에 비하여 심한 우울과 불안 증상을 경험하고 있으며, 비적응적인 수동적 대처방식을 사용함으로써 이런 부정적 감정 경험의 정도가 심해짐을 알 수 있었다. 이는 조혈모세포 이식을 받는 환자의 스트레스 대처방석에 대한 적절한 평가와 정신과적 개입의 필요성을 시사한다고 하겠다.

• 수산해양기술연구
• /
• 제32권3호
• /
• pp.273-285
• /
• 1996

Generally, the flow around the ship's hull often appear 3-dimensional separation at the bow and stem, and the vortex by this flow affects greatly the resistance propulsive efficiency and maneuverability of the ship. This study is compared the calculated result based on the streamlines calculation method with the experimental result by oil fIlm method to analyze the patterns and characteristics of the flows around the ship's hull of chine form. DTMB Series 62, 4667-1 vessel was selected as a sample ship of chine form and model ship was painted with the mixture of oil-color, paint, and poly wax at the surface of the vessel and tested in the model towing tank of Inha University. The results obtained in this study are listed briefly as follows ; 1. For the single chine form, after $4\frac{1}{2}$ station the streamlines are crossed at the chine line and the streamlines are converted to the vortex follow the chine line. 2. For the single chine form, the vortex appered increases and severely in and arround the stern. 3. The approximate streamlines for the ship's hull of chine form can be assigned by the use of the Watanabe's basic transformation formular.