• 한국발생생물학회지:발생과생식
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• 제19권3호
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• pp.119-126
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• 2015

The suitable feeder cell layer is important for culture of embryonic stem (ES) cells. In this study, we investigated the effect of two kinds of the feeder cell, MEF cells and STO cells, layer to mouse ES (mES) cell culture for maintenance of stemness. We compare the colony formations, alkaline phosphatase (AP) activities, expression of pluripotency marker genes and proteins of D3 cell colonies cultured on MEF feeder cell layer (D3/MEF) or STO cell layers (D3/STO) compared to feeder free condition (D3/-) as a control group. Although there were no differences to colony formations and AP activities, interestingly, the transcripts level of pluripotency marker genes, Pou5f1 and Nanog were highly expressed in D3/MEF (79 and 93) than D3/STO (61and 77) or D3/- (65 and 81). Also, pluripotency marker proteins, NANOG and SOX-2, were more synthesized in D3/MEF ($72.8{\pm}7.69$ and $81.2{\pm}3.56$) than D3/STO ($32.0{\pm}4.30$ and $56.0{\pm}4.90$) or D3/- ($55.0{\pm}4.64$ and $62.0{\pm}6.20$). These results suggest that MEF feeder cell layer is more suitable to mES cell culture.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제16권8호
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• pp.1102-1107
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• 2003

The ES cell can provide a useful system for studying differentiation and development in vitro and a powerful tool for producing transgenic animalds. To investigate the culture condition of chicken embryonic stem (CES) cells which can retain their multipotentiality or totipotency, three kinds of feeder layer cells, SNL cells, primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells, were used as the feeder cells in media of DMEM supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and stem cell factor (SCF) for co-culture with blastoderm cells from stage X embryos of chicken. The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The results showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The typical CES cells clone shape revealed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong alkaline phosphatase (AKP) reactive cells were observed in the fourth passage cells. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2% (22/269) survived to hatching, 5 feather chimeras had been produced. This suggests that an effective culture system established in this study can promote the growth of CES cells and maintain them in the state of undifferentiated and development, which lays a solid foundation for the application of CES cells and may provide an alternative tool for genetic modification of chickens.

• 한국수정란이식학회지
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• 제26권2호
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• pp.111-115
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• 2011

In general, zona pellucida (ZP) of the blastocyst has to be removed first, then either isolated the inner cell mass (ICM) or ZP-removed whole blastocyst, which is then cultured on the feeder layer to induce ICM outgrowth for the generation of embryonic stem cells (ESC). However, it is unclear whether ICM isolation before seeding on feeder layer is beneficial or not because the interaction between ICM and trophoblasts may affect cellular growth and/or pluripotency during the culture on the feeder. In the present study, two ZP removal methods (mechanically by splitting with a 28-gauge needle versus chemically by the treatment of acid-Tyrode's solution) and two ICM isolation methods (ZP-free whole blastocyst seeding versus mechanical isolation of ICM) were evaluated for the efficient isolation and culture of putative parthenogenetic bovine ESC. The number of maintained outgrown colonies was counted in each experimental group. As the result, mechanical removal of ZP with a needle and followed by whole ZP-free blastocyst seeding on feeder cells tended to attach more on the feeder layer and resulted in more outgrown colonies with its simple and less time-costing benefits. Currently we are generating ESC lines in HanWoo cattle by using this method for initial outgrowth of the parthenogenetic bovine blastocysts.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.261-271
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• 2004

Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at $37^{circ}C$ in a 5% $CO_2$ in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.

• 한국정보통신학회:학술대회논문집
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• 한국해양정보통신학회 1999년도 추계종합학술대회
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• pp.116-119
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• 1999

This paper presents a feeder waveguide array of slot array antennas for Direct Broadcasting from Satellite(DBS). The feed structure consists of a single waveguide placed on the same layer as radiating waveguide and is characterized by the unit divider, called a $\pi$-Junction. This K-function with an inductive wall splits part of the power into two branch waveguide through one coupling window, and can excite densely arrayed waveguide at equal phase and amplitude. The power dividing characteristics of a cascade of $\pi$ -functions are analyzed by Galerkin's method of moments. From the optimum simulation results based on the feeder waveguide using $\pi$ -Junction at 3.95GHz, we obtained the scattering matrices of the feeder divided power at 11.85CHz.

• 한국전자파학회논문지
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• 제12권2호
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• pp.257-267
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• 2001

본 논문은 유도성 벽을 이용한 $\pi$분기형 일층구조 급전도파관 어레이 설계에 대하여 기술하고 있다. 이 구조는 복사도파관과 급전도파관을 같은 층에 위치시킴으로써 전체 구조를 일층구조로 만들었으며, 유도성 벽을 이용함으로써 하나의 도파관 창으로부터 복사도파관으로 급전부의 전력분배를 동위상. 동진폭으로 분배되도록 설계하였다. Galerkin's 모멘트법을 이용하여 유동성 벽을 포함한 다단성 급전도파관을 엄밀하게 해석하였고, 설계에 있어서 전송선로형 등가회로 기념을 이용하여 전력분배비와 반사계수를 구하였다. $\pi$결합분기기 한 단에 대해 시뮬레이션하고 그 값을 토대로 하여 실제 제작을 통해 그 타당성을 입증하였고, 한 단에 대한 설계 방법을 토대로 반복 계산에 의한 설계 주파수 3.95GHz을 중심으로 하는 8-port 어레이 급전구조를 설계하였다.

• 한국발생생물학회지:발생과생식
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• 제12권2호
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• pp.141-149
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• 2008

This study was conducted to develop an efficient cryopreservation method of human embryonic stem (ES) cells using vitrification. In an initial experiment, sub-clumps of human ES cells (CHA-hES3 and CHA-hES4) were vitrified using grids after incubation with STO feeder cells for 1 or 16 h (Groups 1-1 and 1-2, respectively). After storage for $2{\sim}4$ months, thawed clumps were re-plated on a fresh feeder layer. The survival rates of warmed CHA-hES3 and CHA-hES4 cells of Group 1-2 were significantly higher than those of the corresponding Group 1-1 cells. In the second experiment, human ES cells were vitrified after incubation with feeder or feeder-conditioned medium (Groups 2-1 to -7). Relative mRNA expression of BM proteins and survival rates were increased following incubation of ES cells with fresh feeder cells for 16 h. In conclusion, increasing of tight adhesion between ES cells by extended incubation with feeder could reduce cryoinjury after vitrifying/warming.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.77-77
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• 2003

Coculture of HSC with bone marrow-derived mesenchymal stem cells (BM-MSCs) is one of used methods to increase cell numbers before transplant to the patients. However, because of difficulties to purify HSCs after coculture with BM-MSCs, it needs to develop a method to overcome the problem. In the present study, we have examined whether a culture insert placed over a feeder layer might support the expansion of HSCs within the insert. $CD34^+/ $ cells isolated from the umbilical cord blood by using midiMACS were divided into three groups. A group of 1 $\times$ $10^5$ cells were grown on a culture insert without feeder layer (Direct). The same number of HSCs was directly cocultured with BM-MSCs (Contact). The third group was placed onto an insert below which BM-MSCs were grown (Insert). To distinguish feeder cells from HSCs, BM-MSCs was pre-labeled fluorescently with PKH26 and 1 $\times$ $10^5$ cells were seeded in the culture dishes. After culture for 13 days, the expansion factor (x) of HSCs that were grown without feeder layer (Direct) was $26.6 \pm 8.4.$ In contrast, the number of HSCs directly cocultured with feeder layer was 59.6 $\pm$ 0.5 and that of HSCs cultured onto an insert was $46.9 \pm 8.4.$ The percentage of BM-MSCs cells remained being fluorescent was $97.9 \pm 0.3%$ after culture. Immune-phenotypically large proportion of cultured cells were founded to be differentiated into myeloid/monocyte progenitor cells. The ability of BM-MSCs, fetal lung, cartilage and brain tissue cells to support ex vivo expansion of HSCs was also examined using the insert. After 11 days of coculture with each of these cells, the expansion factor of HSCs was 15.0, 39.0, 32.0 and 24.0, respectively. Based upon these observations, it is concluded that the coculture method using insert is very effective to support ex vivo expansion of HSCs and to eliminate the contamination of other cells used to coculture wth HSCs.

• 한국축산시설환경학회지
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• 제14권1호
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• pp.53-60
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• 2008

최근 한국의 국민 소득 증대로 인해 육류 및 계란의 소비량이 점차적으로 증가하여, 산란계사의 규모가 대형화됨에 따라 기계화되었고, 기계 시설의 소음/진동이 계사 내 가축의 생산능력에 직 간접적으로 영향을 미치는 영향에 대한 선행연구는 일부분에 국한되어 있다. 이에 본 연구에서는 계사내 기계장치 중 진동발생량이 가장 큰 호퍼식급이기의 진동으로 인한 공진을 미연에 방지하고자 진동모드 및 진동발생량을 계측 분석하였다. 이때 축은 급이기의 구동방향(X축), 케이지 방향(Y축) 및 급이기의 상하방향(Z축)으로 설정하였으며, 사료 충전율 0, 25, 50, 75, 100%인 지점에서 매달아서 계측하는 타격여진법을 응용하여 진동모드를 계측하였다. 획득한 데이터를 분석프로그램을 이용하여 분석한 결과, $30{\sim}500Hz$의 호퍼식 급이기 고유주파수를 구명하였고, 특히 170 Hz 이하의 낮은 주파수대에서는 전달율비가 대부분 높은 수준이었다. 이를 근거로 하여 30, 100, 170 Hz대의 호퍼식급이기 진동 시뮬레이션 분석을 실시하여 취약지점에 대한 분석을 실시하여 진동발생량 계측 지점으로 지정, 계측 분석을 실시하였다. 진동모드와 진동발생량 분석결과를 근거로 하여 호퍼식급이기가 산란계 산란율을 영향을 주는 수준의 진동을 발생시킴을 구명하였고, 각 취약지점에 대한 분석을 통하여 각 축별로 공진 및 제진/방진을 위한 방안 및 장치를 제시하였다.