• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.40-46
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• 2001

The use of pluripotent stem cells has tremendous advantages for various purposes but these cell lines with proven germ-line transmission have been completely established only in the mouse. Embryonic germ (EG) cell lines are also pluripotent and undifferentiated stem cells established from primordial germ cells (PGCs). This study was conducted to establish and characterize the chicken EG cells derived from gonadal primordial germ cells. We isolated gonadal PGCs from 5.5-day-old (stage 28) White leghorn (WL) embryos and established chicken EG cells lines with EG culture medium supplemented with human stem cell factor (hSCF), murine leukemia inhibitory factor (mLIF), bovine basic fibroblast growth factor (bFGF), human interleukin-11 (hIL-11), and human insulin-like growth factor-I (hIGF-I). These cells grew continuously for 4 months (10 passages) on a feeder layer of mitotically active chicken embryonic fibroblasts. These cells were characterized by screening with the Periodic acid-Shiff＇s reaction, anti-SSEA-1 antibody, and a proliferation assay after several passages. As the results, the chicken EG cells maintained characteristics of undifferentiated stem cells as well as that of gonadal PGCs. When cultured in suspension, the chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types when re-seeded onto culture dish. The chicken EG cells were injected into blastodermal layer at stage X and dorsal aorta of recipient embryo at stage 14 (incubation of 53hrs) and produced chimeric chickens with various differentiated tissues derived from the EG cells. The germline chimeras were also successfully induced by using EG cells. Thus, Chicken EG cells will be useful for the production of transgenic chickena and for studies of germ cell differentiation and genomic imprinting.

• Molecules and Cells
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• 제25권3호
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• pp.358-367
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• 2008

The embryonic germ cell (EGCs) of mice is a kind of pluripotent stem cell that can be generated from pre- and post-migratory primordial germ cells (PGCs). Most previous studies on DNA methylation of EGCs were restricted to 12.5 days post coitum (dpc). This study was designed to establish and characterize murine EGC lines from migrated PGCs as late as 13.5 dpc and to estimate the degrees of methylation of their imprinted genes as well as of the non-imprinted locus, Oct4, using an accurate and quantitative method of measurement. We established five independent EGC lines from post migratory PGCs of 11.5-13.5 dpc from C57BL/6 ${\times}$ DBA/2 F1 hybrid mouse fetuses. All the EGCs exhibited the typical features of pluripotent cells including hypomethylation of the Oct4 regulatory region. We examined the methylation status of three imprinted genes; Igf2, Igf2r and H19 in the five EGC lines using bisulfite genomic sequencing analysis. Igf2r was almost unmethylated in all the EGC lines irrespective of the their sex and stage of isolation; Igf2 and H19 were more methylated than Igf2r, especially in male EGCs. Moreover, EGCs derived at 13.5 dpc exhibited higher levels of DNA methylation than those from earlier stages. These results suggest that in vitro derived EGCs acquire different epigenotypes from their parental in vivo migratory PGCs, and that sex-specific de novo methylation occurs in the Igf2 and H19 genes of EGCs.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.791-797
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• 2018

Objective: Trimethylation of histone 3 (H3) at 4th lysine N-termini (H3K4me3) in gene promoter region was the universal marker of active genes specific to cell lineage. On the contrary, coexistence of trimethylation at 27th lysine (H3K27me3) in the same loci-the bivalent H3K4m3/H3K27me3 was known to suspend the gene transcription in germ cells, and could also be inherited to the developed stem cell. In galline species, throughout example of H3K4m3 and H3K27me3 ChIP-seq analysis was still not provided. We therefore designed and demonstrated such procedures using ChIP-seq and mRNA-seq data of chicken follicular mesenchymal cells and male germ cells. Methods: Analytical workflow was designed and provided in this study. ChIP-seq and RNA-seq datasets of follicular mesenchymal cells and male germ cells were acquired and properly preprocessed. Peak calling by Model-based analysis of ChIP-seq 2 was performed to identify H3K4m3 or H3K27me3 enriched regions ($Fold-change{\geq}2$, $FDR{\leq}0.01$) in gene promoter regions. Integrative genomics viewer was utilized for cellular retinoic acid binding protein 1 (CRABP1), growth differentiation factor 10 (GDF10), and gremlin 1 (GREM1) gene explorations. Results: The acquired results indicated that follicular mesenchymal cells and germ cells shared several unique gene promoter regions enriched with H3K4me3 (5,704 peaks) and also unique regions of bivalent H3K4m3/H3K27me3 shared between all cell types and germ cells (1,909 peaks). Subsequent observation of follicular mesenchyme-specific genes-CRABP1, GDF10, and GREM1 correctly revealed vigorous transcriptions of these genes in follicular mesenchymal cells. As expected, bivalent H3K4m3/H3K27me3 pattern was manifested in gene promoter regions of germ cells, and thus suspended their transcriptions. Conclusion: According the results, an example of chicken H3K4m3/H3K27me3 ChIP-seq data analysis was successfully demonstrated in this study. Hopefully, the provided methodology should hereby be useful for galline ChIP-seq data analysis in the future.

• Journal of Animal Science and Technology
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• 제49권2호
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• pp.183-194
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• 2007

본 연구는 한우에서 이유시 체중과 도체형질들 간의 상관관계를 구명하고자 실시하였다. 조사된 도체 형질은 배최장근 단면적, 등지방두께, 21단계로 구분하여 평가한 근내지방도(근내지방도 I)와 7단계로 구분하여 평가한 근내지방도(근내지방도 II), 그리고 7단계로 구분하여 평가한 육색 등이었다. 유전모수의 추정은 DFREML 방법을 적용하여 실시하였는데 이유시 체중에 대한 통계모형은 동기우 그룹효과(년도－계절－성) 외에 이유시 송아지 일령 및 어미소 일령의 1차식 효과와 2차식 효과를 고정효과로 포함하였고 개체효과를 임의효과로 포함하였다. 도체형질에 대한 통계 모형은 동기우 그룹효과(년도－계절－성) 외에 도축시 일령의 일차식 효과를 고정효과에 포함하였고 개체효과를 임의효과로 포함하였다. 조사된 형질별 유전력 추정치는 이유시 체중이 0.25, 배최장근 단면적이 0.20, 등지방두께가 0.20, 근내지방도Ⅰ이 0.32, 근내지방도 Ⅱ가 0.32 그리고 육색이 0.22였다. 이유시 체중과 배최장근 단면적, 등지방두께, 근내지방도 I, 근내지방도 II 및 육색 간의 유전(표현형) 상관계수는 각각 0.75(0.16), 0.18(0.05), －0.41(－0.09), －0.40(0.11) 및 －0.07(0.05)였다. 본 연구 결과는 이유시 체중이 무거운 방향으로 단형질 선발을 진행할 경우 등심단면적이 넓어지고, 등지방두께가 두꺼워지며 근내지방도가 감소하는 도체를 생산하는 후손집단이 형성될 가능성이 있음을 시사하고 있다.

• 한국발생생물학회지:발생과생식
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• 제15권1호
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• pp.41-52
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• 2011

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.74-76
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• 2001

조류의 경우에는 포유류와 달리 수정란의 성별이 암컷에 의하여 결정된다. 수컷은 동일접합체로 ZZ 염색체를, 암컷의 경우에는 이형접합체로 Z W 염색체를 갖기 때문이다. 현재까지 조류에 있어서 염색체 분석 등에 의한 암 ·수의 세포 유전학적인 특성은 많은 연구가 되어 있으나, 배발달 초기의 원시생식세포 등에 대해서는 많은 연구가 진행되어 있지 않다. 따라서 본 연구는 암컷의 원시생식세포를 분리하여 숫컷의 초기 배자에 주입함으로써 수용체 배자의 원시생식기내로 이동이 가능한지를 검증하였으며, 또한 수컷의 원시생식기내로의 이동 후 정상적으로 분열 및 분화가 가능한지를 초기 배발달 과정에서 확인하였다. 본 연구 결과, 암컷의 원시생식세포는 수컷의 수용체 배자에 재주입시 정상적인 원시생식기내로의 이동 능력을 보여주었으며, 분열 ·분화함을 알 수있었다.

• 한국식품과학회지
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• 제51권2호
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• pp.97-102
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• 2019

밀 배아를 셀룰레이즈 활성이 있는 Celluclast 1.5L을 이용하여 다양한 시간 및 온도의 반응에서 생성되는 유기산(주석산, 젖산, 초산, 호박산)과 유리당(포도당, 설탕, 과당) 함량을 분석하였다. 또한 젖산, 초산, 호박산, 주석산으로 pH 4.5로 조정한 유기산 용액에서 Celluclast 1.5L과 밀 배아를 반응시켜 2,6-DMBQ의 함량에 미치는 영향을 평가하였다. 밀 배아에 대한 효소 반응 시간 및 온도가 증가함에 따라 주석산, 젖산, 초산, 호박산과 이들의 총유기산 함량은 증가하였다. 밀 배아 효소 반응 추출액의 설탕 농도는 효소 반응 시간이 증가할수록 지속적으로 감소한 반면에, 과당과 포도당의 농도는 효소 반응 시간이 길어짐에 따라 증가하였다. 유기산을 첨가한 Celluclast 1.5L 효소 추출 용액으로 반응시킨 밀 배아는 반응시간이 증가할수록 2,6-DMBQ 함량이 증가하는 경향을 보였지만, 대조군 대비 향상된 수치를 보이지 않았다. 향후 효소 처리 밀 배아 추출물을 활용한 다양한 생리활성에 대한 효능 평가와 더불어 효소 처리 밀 배아 추출물의 식품 소재화에 대한 연구가 필요할 것으로 보인다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 2006년도 제61회 한국생물과학협회 정기학술대회
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• pp.9-9
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• 2006

• Journal of Nutrition and Health
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• 제53권2호
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• pp.99-110
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• 2020

본 연구에서는 고기능성 쌀인 큰눈자미 배아의 phytosterols 함량 및 in vitro 항산화 활성을 평가하고, 성숙기 흰쥐에게 보충 급여한 당질 대사, 항산화 활성 및 일부 cytokines 개선 효과 여부를 검증하였다. Phytosterols 분석에서 NG보다 KG의 campesterol과 stigmasterol의 함량이 유의하게 높았다. NG에 비해 KG가 DPPH radical 소거 활성, 환원력 및 ABTS radical 소거능 측정에서 유의적으로 높은 값을 나타내었다. 실험동물은 각 10마리씩 3군으로 나누어 일반 식이를 급여하는 NC군, 일반현미배아 3%를 첨가하는 NG3군, 큰눈자미배아 3%를 첨가하는 KG3군으로 나누어 사육하였다. 그 결과 KG3군에서 체중증가량, 신장주위 및 총 지방량이 유의하게 감소하였다. 당질 대사에서 실험군들 간에 glucose, 인슐린, C-peptide 및 HOMA-IR의 수준이 유의적인 차이가 나타나지 않았다. KG3군에서 혈중 TNF-α 수준이 NG3에 비해 유의적으로 감소하고, SOD 활성이 유의하게 증가하였으며, leptin, AOPP 및 IL-6 수준이 감소하는 경향을 나타내었다. 이상의 결과로부터 큰눈자미 배아는 높은 함량의 phytosterols과, 우수한 in vitro 항산화활성, 그리고 in vivo 실험에서 일부의 cytokine 개선 및 항산화에 긍정적인 효과가 있음을 제시하였고 향후 더 많은 생리활성물질 분석, 대사 지표 개선, 작용 기전 규명 등 세부적인 연구가 필요할 것으로 사료된다.

• Journal of Biosystems Engineering
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• 제29권2호
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• pp.121-130
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• 2004

This study was carried out to develop a miller to produce white embryo rice with functional nutrients by improving the conventional vertical miller. The effects of rice moisture content and the shaft revolution speed of the miller on germ(embryo) adherence rate, whiteness, broken rice rate, and cracked rice rate were investigated. Also, the effect of the mesh size of emery stones on the germ adherence rate was investigated. The vertical prototype miller was improved with the increasement of about 42% in producing white embryo rice at proper conditions(shaft revolution speed of 900 rpm, emery stones of mesh ＃50, processing capacity of 2.3t/h, zero outlet resistance, rice moisture content of 16.2%). The results were as follows: 1. The germ adherence rate of white rice was significantly influenced by the moisture content of brown rice. The germ adherence rate of white rice decreased rapidly with the increase of the moisture content of brown rice. When brown rice with moisture content of 13.2%, 14.5%, 15.2%, 15.4% was milled by the prototype with emery stones of mesh ＃35 and shaft speed of 900(1,100) rpm, rpm adherence rate of milled rice was 76.2%(70%), 69.2%(66%), 45.9(38%), 13.0(9%), respectively. 2. The whiteness of white rice milled by the prototype with emery stones of mesh ＃35 and shaft speed of 1,100(900)rpm increased from 27(23) to about 40, respectively, as the moisture content of brown rice increased from 13.2% to 17.2%. 3. The rate of broken rice of white rice milled at 900rpm decreased by 0.6∼1.0% compared with that at 1,100rpm when the moisture content of brown rice was less than 15.2%. 4. The germ adherence rate was increased by 10.3% and 11.0%, respectively when brown rice with moisture content of 16.2% and 15.5% was milled by the prototype miller with shaft speed of 900rpm and emery stones of mesh ＃50 instead of mesh ＃35. 5. Considering the germ adherence rate, broken rice rate, and whiteness of milled rice, the proper milling conditions of the prototype miller for producing embryo rice were the moisture content of about 15%, the processing capacity of 2.3t/h and minimum outlet resistance of 0Nㆍm with shaft speed of 900rpm and emery stones of mesh ＃50.