• Reproductive and Developmental Biology
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• 제36권4호
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• pp.291-294
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• 2012

Embryonic stem cell classically cultured on feeder layer with FBS contained ES medium. Feeder-free mouse ES cell culture systems are essential to avoid the possible contamination of nonES cells. First we determined the difference between ES cell and MEF by Oct4 population. We demonstrate to culture and to induce differentiation on feeder free condition using a commercially available mouse ES cell lines.

• Journal of Plant Biotechnology
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• 제30권3호
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• pp.257-261
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• 2003

This experiment was carried out to investigate the effects of plant growth regulators on the organogenesis from the tissue culture of Arabidopsis thaliana stem, and the origin of the callus development. When the stem segments were cultured on medium with 2mg/L IAA or NAA, adventitious roots and trichomes were differentiated after 11 days of culture. Callus vigorously formed on medium with 2/L2,4 after 7 days of culture, but adventitious roots and trichomes were not differentiated from callus after 10 days of culture. This results suggesting that picloram is very effective auxin for the callus formation and organogenesis. Callus weakly formed on 0.05mg/L kinetin, and formed on combination of auxins(2mg/L) with 0.05mg/L kinetin. But the effect of combination of auxins and kinetin the callus formation was less than 2,4-D or picloram alone. A histological examination of callus formed on picloram showed that phloram showed that phloem parenchyma cells were divided and enlarged after 2 days of culture. Cortex parenchyma cells were divided and meristematic nodules were developed from these cells after 4 days of culture. Finally, callus formed on outside of cortex and epidermis by division of meristematic nodules after 7 days of culture.

• Molecules and Cells
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• 제41권7호
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• pp.631-638
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• 2018

Spermatogonial stem cells (SSCs) derived from mouse testis are unipotent in regard of spermatogenesis. Our previous study demonstrated that SSCs can be fully reprogrammed into pluripotent stem cells, so called germline-derived pluripotent stem cells (gPS cells), on feeder cells (mouse embryonic fibroblasts), which supports SSC proliferation and induction of pluripotency. Because of an uncontrollable microenvironment caused by interactions with feeder cells, feeder-based SSC reprogramming is not suitable for elucidation of the self-reprogramming mechanism by which SSCs are converted into pluripotent stem cells. Recently, we have established a Matrigel-based SSC expansion culture system that allows longterm SSC proliferation without mouse embryonic fibroblast support. In this study, we developed a new feeder-free SSC self-reprogramming protocol based on the Matrigel-based culture system. The gPS cells generated using a feeder-free reprogramming system showed pluripotency at the molecular and cellular levels. The differentiation potential of gPS cells was confirmed in vitro and in vivo. Our study shows for the first time that the induction of SSC pluripotency can be achieved without feeder cells. The newly developed feeder-free self-reprogramming system could be a useful tool to reveal the mechanism by which unipotent cells are self-reprogrammed into pluripotent stem cells.

• KSBB Journal
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• 제31권2호
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• pp.120-125
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• 2016

Two-dimensional cultivation is typically used for cell growth, but the method reduces the characteristics of chondrocytes and stem cells, and limits culture area. Therefore, development of three-dimensional culture method is needed to mimic in vivo environment, improve quality of cells and scale-up efficiently. Improving proliferation and chondrogenesis is available by co-culture of chondrocytes and mesenchymal stem cells (MSCs) that leads to interaction between two kinds of cells. However, the co-culture has problems that permeability of sphere diminishes as aggregate size increased and ratio of two kinds of cells composing each spheres is different. In this work, co-cultivation method using controlled sphere composed of chondrocytes and MSCs was established and enhanced chondrogenesis. Periosteum-derived progenitor cells (PDPCs) that are appropriate for cell therapy source of articular cartilage were used as MSCs. Controlled spheres were formed in the hanging-drop plates and shifted for being induced chondrogenesis in 35-mm non-adhesive culture dishes at a rotation rate of 60 rpm. After inducing chondrogenesis, gene expressions related with chondrogenesis were found to be improved and it was apparent that the utilization of controlled spheres promoted chondrogenesis. As a result, available numbers of cells per unit area were increased and chondrogenic differentiation ability was improved compared to typical two-dimensional culture. This approach shows the potential in cartilage regeneration as it can provide sufficient numbers of chondrocytes.

• 한국수정란이식학회지
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• 제29권3호
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• pp.221-228
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• 2014

Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preserving eternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes of specific human diseases, there were no reports about precise definition of niche conditions promoting proliferation and maintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenance of undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in different composition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors, and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of $4{\times}10^5$ pSSCs on mitotically in activated $2{\times}10^5$ STO cells in the mouse embryonic stem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF) was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCs in the following researches.

• International Journal of Oral Biology
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• 제43권2호
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• pp.77-82
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• 2018

The mesenchymal stem cells (MSCs) that reside in dental tissues hold a great potential for future applications in regenerative dentistry. In this study, we used human dental pulp cells, isolated from the molars (DPCs), in order to establish the organoid culture. DPCs were established after growing pulp cells in an MSC expansion media (MSC-EM). DPCs were subjected to organoid growth media (OGM) in comparison with human dental pulp stem cells (DPSCs). Inside the extracellular matrix in the OGM, the DPCs and DPSCs readily formed vessel-like structures, which were not observed in the MSC-EM. Immunocytochemistry analysis and flow cytometry analysis showed the elevated expression of CD31 in the DPCs and DPSCs cultured in the OGM. These results suggest endothelial cell-prone differentiation of the DPCs and DPSCs in organoid culture condition.

• Asian-Australasian Journal of Animal Sciences
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• 제20권10호
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• pp.1510-1516
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• 2007

These experiments were carried out to optimize the parameters of electrical activation, methods of parthenogenetic activation and embryo culture in vitro and meanwhile to isolate embryonic stem cells-like (ESCs) derived from porcine parthenogenetic blastocysts (pPBs). These results showed that, as the electric field strength increased from 1.0 to 2.7 kV/cm, the cleavage rate of parthenogenetic embryos increased gradually but the rate of oocyte lysis was significantly increased when using 2.7 kV/cm field strength. The rate of cleavage in 2.2 and 2.7 kV/cm groups was significantly increased in comparison with that of the 1.0 kV/cm group. A voltage field strength of 2.2 kV/cm DC was used to investigate blastocyst development following activation with a single pulse of 30 or $60-{\mu}sec$ pulse duration. The optimum pulse duration was 30-${\mu}sec$, with a blastocyst rate of 20.7%. Multiple pulses were inferior to a single pulse for blastocyst yield (8.0% vs. 29.9) (p<0.05). For porcine oocyte parthenogenetic activation methods, the rates of cleavage (79.0% vs. 59.8%) and blastocysts (19.4% vs. 3.4%) were significantly increased in electrical activation in contrast to chemical activation with ionomycin/6-DMAP (p<0.05). Rates of cleavage and blastocyst formation in NCSU-23 and PZM-3 embryo media were higher than those of G1.3/G2.3 serial culture media, but there was no significant difference among the three groups. The total cell number of blastocysts in PZM-3 embryo culture media containing $5{\mu}g/ml$ insulin was significantly higher than that of the control (no insulin) ($44.3{\pm}9.1$ vs. $33.9{\pm}11.7$). For isolation of PESCs-like, the rates of porcine blastocysts attached to feeder layers and ICM colony formation in Method B (nude embryo culture) were better than those in Method A (intact embryo culture).

• 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.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.54-57
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• 2006

• International Journal of Advanced Culture Technology
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• 제8권3호
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• pp.280-284
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• 2020

In the rapidly changing science and technology environment, an in-depth understanding of users of STEM information is an essential factor in designing a user-centered information system. The purpose of this study is to investigate and analyze in-depth the behaviors and needs of users who use STEM information. In this study, the needs of users for STEM information and STEM information sites are dealt with in depth using the FGI qualitative method. In addition, it includes the analysis results of grouping of similar sites according to various aspects of use of STEM information sites. As a result of grouping similar sites based on awareness and level of use,, they were grouped by domestic-international, paid-free, integrated-specific fields. As a result of grouping similar sites according to the purpose of use, they were grouped by domestic and international papers, research reports, and patents. As a result of grouping similar sites according to usage attributes, they were grouped by diversity, reliability, and specialization. As for the positions of similar sites perceived by users, Science Direct and PubMed showed high specialization and high quality, Google Scholar showed integration and popularity, and RISS showed four attributes evenly. Suggestions for information system design are discussed.