http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Han, Na Rae,Baek, Song,Lee, Yongjin,Lee, Joohyeong,Yun, Jung Im,Lee, Eunsong,Lee, Seung Tae The Korean Society of Animal Reproduction and Biot 2020 한국동물생명공학회지 Vol.35 No.1
The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.
Han, Na Rae,Park, Hye Jin,Lee, Hyun,Yun, Jung Im,Choi, Kimyung,Lee, Eunsong,Lee, Seung Tae The Korean Society of Embryo Transfer 2018 한국동물생명공학회지 Vol.33 No.4
To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.
Han, Mi Na,Byeon, Hyeon Seop,Han, Seong Tae,Jang, Rae Hoon,Kim, Chang Seop,Choi, Seok Hwa The Korean Society of Veterinary Service 2018 韓國家畜衛生學會誌 Vol.41 No.4
Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.
Han, Na Rae,Lee, Hyun,Yun, Jung Im,Kim, Choonghyo,Hwang, Jae Yeon,Park, Kyu Hyun,Lee, Seung Tae The Korean Society of Embryo Transfer 2017 한국동물생명공학회지 Vol.32 No.2
Mesenchymal stem cells (MSCs) have been considered an alternative source of neuronal lineage cells, which are difficult to isolate from brain and expand in vitro. Previous studies have reported that MSCs expressing Nestin ($Nestin^+$ MSCs), a neuronal stem/progenitor cell marker, exhibit increased transcriptional levels of neural development-related genes, indicating that $Nestin^+$ MSCs may exert potential with neurogenic differentiation. Accordingly, we investigated the effects of the presence of $Nestin^+$ MSCs in bone-marrow-derived primary cells (BMPCs) on enhanced neurogenic differentiation of BMPCs by identifying the presence of $Nestin^+$ MSCs in uncultured and cultured BMPCs. The percentage of $Nestin^+$ MSCs in BMPCs was measured per passage by double staining with Nestin and CD90, an MSC marker. The efficiency of neurogenic differentiation was compared among passages, revealing the highest and lowest yields of $Nestin^+$ MSCs. The presence of $Nestin^+$ MSCs was identified in BMPCs before in vitro culture, and the highest and lowest percentages of $Nestin^+$ MSCs in BMPCs was observed at the third (P3) and fifth passages (P5). Moreover, significantly the higher efficiency of differentiation into neurons, oligodendrocyte precursor cells and astrocytes was detected in BMPCs at P3, compared with P5. In conclusion, these results demonstrate that neurogenic differentiation can be enhanced by increasing the proportion of $Nestin^+$ MSCs in cultured BMPCs.
Na Rae Han,Song Baek,Yongjin Lee,Joohyeong Lee,Jung Im Yun,Eunsong Lee,Seung Tae Lee 한국동물생명공학회(구 한국동물번식학회) 2020 Journal of Animal Reproduction and Biotechnology Vol.35 No.1
The establishment of porcine embryonic stem cells (ESCs) from porcine somatic cell nuclear transfer (SCNT) blastocysts is influenced by in vitro culture day of porcine reconstructed embryo and feeder cell type. Therefore, the objective of the present study was to determine the optimal in vitro culture period for reconstructed porcine SCNT embryos and mouse embryonic fibroblast (MEF) feeder cell type for enhancing colony formation efficiency from the inner cell mass (ICM) of porcine SCNT blastocysts and their outgrowth. As the results, porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days showed significantly increased efficiency in the formation of colonies, compared to those for 7 days. Moreover, MEF feeder cells derived from outbred ICR mice showed numerically the highest efficiency of colony formation in blastocysts produced through in vitro culture of porcine SCNT embryos for 8 days and porcine ESCs with typical ESC morphology were maintained more successfully over Passage 2 on outbred ICR mice-derived MEF feeder cells than on MEF feeder cells derived from inbred C57BL/6 and hybrid B6CBAF1 mice. Overall, the harmonization of porcine SCNT blastocysts produced through in vitro culture of the reconstructed embryos for 8 days and MEF feeder cells derived from outbred ICR mice will greatly contribute to the successful establishment of ESCs derived from porcine SCNT blastocysts.
Identification of a Technique Sorting Effectively Spermatogonial Stem Cells from Mouse Testes
Na Rae Han,Hye Jin Park,Hyun Lee,Seung Tae Lee 한국동물생명공학회(구 한국동물번식학회) 2018 발생공학 국제심포지엄 및 학술대회 Vol.2018 No.06
Spermatogonial stem cells (SSCs) can produce sperms transferring genetic information into the next generation in seminiferous tubule of testes. Accordingly, an efficient isolation technique of SSCs with extremely low numbers from testes is required for successful downstream researches related to maintenance, differentiation and cryopreservation of SSCs. To date, a variety of isolation techniques in a variety of species have been used for retrieving SSCs from testicular tissue. However, comparison of their efficiency has been not revealed clearly. Accordingly, among isolation methods described previously, we tried to elucidate a technique showing the best isolation efficiency in the retrieval of SSCs from testes derived from mice. For these, SSCs were isolated from mouse testis as follows: differential plating (DP), EpCAM, Thy1 or GFRα1 antibody-based magnetic-activating cell sorting (MACS) post-DP, EpCAM, Thy1 or GFR α1 antibody-based MACS, EpCAM, Thy1 or GFRα1 antibody-based MACS post-MACS based on GFRα1 antibody (double MACS for positive selection), and CD34 or α-SMA antibody-based MACS post-MACS based on GFRα1 antibody (double MACS for negative selection). Subsequently, SSCs isolated from each method were stained by alkaline phosphatase (AP) staining and percentage of AP positive SSCs was compared to find an optimal method among isolation method candidates. As the results, SSCs isolated by DP for 8 h showed numerically the highest percentage of AP positive SSCs. In case of MACS post-DP, SSCs isolated by MACS based on Thy1 antibody post-DP (MACSThy1 post-DP) revealed numerically higher percentage of AP positive cells than those on EpCAM and GFRα1 antibodies post-DP. Moreover, numerically the highest percentage of AP positive SSCs was detected when SSCs were isolated from mouse testis by MACS based on GFRα1 (MACSGFRα1) compared to EpCAM and Thy1 antibodies. In case of double MACS for positive selection, the usage of EpCAM antibody in the second MACS post-GFRα1 antibody-based MACS (double MACSGFRα1/EpCAM) showed numerically the highest percentage of AP positive SSCs compared to Thy1 and GFRα1 antibodies. On the other hands, after GFRα1 antibody-based MACS sorting, the usage of α-SMA antibody in the second MACS (double MACSGFRα1/α-SMA) for negative selection showed numerically higher percentage of AP positive SSCs than CD34 antibody. The subsequent comparison of isolation efficiency derived from each method demonstrated that MACSGFRα1 and double MACSGFRα1/EpCAM resulted in significantly the best isolation efficiency. Accordingly, we could elucidate that MACSGFRα1 and double MACSGFRα1/EpCAM were techniques sorting effectively SSCs from mouse testes.
Na Rae Han,Hyun Lee,Jung Im Yun,Choonghyo Kim,Jae Yeon Hwang,Kyu Hyun Park,Seung Tae Lee 한국수정란이식학회 2017 한국동물생명공학회지 Vol.32 No.2
Mesenchymal stem cells (MSCs) have been considered an alternative source of neuronal lineage cells, which are difficult to isolate from brain and expand in vitro. Previous studies have reported that MSCs expressing Nestin (Nestin+ MSCs), a neuronal stem/progenitor cell marker, exhibit increased transcriptional levels of neural development-related genes, indicating that Nestin+ MSCs may exert potential with neurogenic differentiation. Accordingly, we investigated the effects of the presence of Nestin+ MSCs in bone-marrow-derived primary cells (BMPCs) on enhanced neurogenic differentiation of BMPCs by identifying the presence of Nestin+ MSCs in uncultured and cultured BMPCs. The percentage of Nestin+ MSCs in BMPCs was measured per passage by double staining with Nestin and CD90, an MSC marker. The efficiency of neurogenic differentiation was compared among passages, revealing the highest and lowest yields of Nestin+ MSCs. The presence of Nestin+ MSCs was identified in BMPCs before in vitro culture, and the highest and lowest percentages of Nestin+ MSCs in BMPCs was observed at the third (P3) and fifth passages (P5). Moreover, significantly the higher efficiency of differentiation into neurons, oligodendrocyte precursor cells and astrocytes was detected in BMPCs at P3, compared with P5. In conclusion, these results demonstrate that neurogenic differentiation can be enhanced by increasing the proportion of Nestin+ MSCs in cultured BMPCs.