• 제목/요약/키워드: Human Mouse

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Induced Pluripotent Stem Cell Generation using Nonviral Vector

  • Park, Si-Jun;Shin, Mi-Jung;Seo, Byoung-Boo;Park, Hum-Dai;Yoon, Du-Hak;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • 제35권4호
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    • pp.449-455
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    • 2011
  • Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by etopic expression of transcription factors. iPS cells are indistinguishable from ES cells in terms of morphology and stem cell marker expression. Moreover, mouse iPS cells give rise to chimeric mice that are competent for germline transmission. However, mice derived from iPS cells often develop tumors. Furthermore, the low efficiency of iPS cell generation is a big disadvantage for mechanistic studies. Nonviral plasmid.based vectors are free of many of the drawbacks that constrain viral vectors. The histone deacetylase inhibitor valproic acid (VPA) has been shown to improve the efficiency of mouse and human iPS cell generation, and vitamin C (Vc) accelerates gene expression changes and establishment of the fully reprogrammed state. The MEK inhibitor PD0325901 (Stemgent) has been shown to increase the efficiency of the reprogramming of human primary fibroblasts into iPS cells. In this report, we described the generation of mouse iPS cells devoid of exogenous DNA by the simple transient transfection of a nonviral vector carrying 2A-peptide-linked reprogramming factors. We used VPA, Vc, and the MEK inhibitor PD0325901 to increase the reprogramming efficiency. The reprogrammed somatic cells expressed pluripotency markers and formed EBs.

Comparison of Three Different Culture Systems for Establishment and Long-Term Culture of Embryonic Stem-like Cells from In Vitro-Produced Bovine Embryos

  • Kim, Daehwan;Park, Sangkyu;Roh, Sangho
    • 한국수정란이식학회지
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    • 제27권3호
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    • pp.189-192
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    • 2012
  • Although embryonic stem cells (ESCs) or ES-like cells are reported from many mammalian species other than the mouse, the culture system for murine ESCs may not be suitable to the other species. Previously many other research groups have modified either human or mouse ESC culture systems for bovine ESC culture. In this study, we compared three different culture mediums consisting of DMEM, ${\alpha}$-MEM or KnockOut$^{TM}$-DMEM (KO), which are modified from human or mouse ESC culture system, for the generation of bovine ESCs. In this study, some pre-requisite events which are important for establishment and long-term propagation of ESCs such as inner cell mass (ICM) attachment on feeder cells, primary colony formation and sustainability after passaging. Once the ICM clumps attached on feeder cells, this was designated as passage 0. In regards to the rate of ICM attachment, ${\alpha}$-MEM was superior to the other systems. For primary colony formation, there was no difference between DMEM and ${\alpha}$-MEM whereas KO showed lower formation rate than the other groups. For passaging, the colonies were split into 2~4 pieces and passed every 5~6 days. From passage 1 to passage 3, DMEM system seemed to be appropriate for maintaining putative bovine ESCs. On the other hand, ${\alpha}$-MEM tended to be more suitable after passage 6. Although ${\alpha}$-MEM support to maintain a ES-like cell progenies to passage 15, all three culture systems which are modified from human or mouse ESC culture media failed to retain the propagation and long-term culture of putative bovine ESCs. Our findings imply that more optimized alternative culture system is required for establishing bovine ESC lines.

암세포 증식에 미치는 인삼과 Vitamin C의 영향 I. 인삼과 Vitamin C 병용에 의한 In Vitro에서 암세포 증식 억제 효과 (Effect of Panax ginseng and Sodium Ascorbate (Vitamin C) Treatment on Cancer Cell Growth I. Synergism of Combined Panax ginseng and Vitamin C Action in vitro)

  • 황우익;손흥수
    • Journal of Ginseng Research
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    • 제13권2호
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    • pp.242-247
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    • 1989
  • The effect of ginseng extract and sodium ascorbate (vitamin C) administered separately or in combination on the some cancer cells cultured in vitro have been examined. Mouse leukemic cells (L1210 and P388), human rectal cancer cells (HRT-18) and human colon cancer cells (HCT-48) were used for the experiment. When given separately, the growth rate for each kind of cancer cell was inhibited In proportion to the concentration of ginseng extract or vitamin C. The inhibitory effect on the growth rate of the cancer cells was stronger in ginseng extract than in vitamin C except for the HCT-48 cells. Based on the cytotoxic activity, combined administration of ginseng extract and vitamin C demonstrated a synergistic inhibition of cancer cell growth. The cytotoxic activities of ginseng extract and vitamin C on the mouse leukemic cells were more sensitive than on human colon cancer cells. And the sensitivity of cytotoxic activity was somewhat different in different cancer cell lines.

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Anticancer effect of mountain ginseng Pharmacopuncture to the nude mouse of lung carcinoma induced by NCI-H460 human non-small cell lung cancer cells

  • Kwon, Ki-Rok
    • 대한약침학회지
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    • 제13권1호
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    • pp.5-14
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    • 2010
  • Objectives : This study was performed to examine the anticancer effect of mountain ginseng Pharmacopuncture(MGP) to the nude mouse of lung carcinoma induced by NCI-H460 human nonsmall lung cancer cells. Methods : Human lung cancer (NCI-H460) cells were cultured and applied to evaluate anti-tumor activity in nude mice. After confirmed tumor growth in mice, MGP was treated per 0.1ml/kg dose to intraperitoneal and intravenous injection everyday for four weeks. And checked the changes in body weights, tumor volume, mean survival time and percent, increase in life span, histo-pathological findings, organ weights, and blood chemistry levels. Results : The results of in vivo study showed that MGP may have potential as growth inhibitor of solid tumor induced NCI-H460 without marked side effects. MGP inhibited dosage-dependently the growth of NCI-H460 cell-transplanted solid tumor compared with the control group. And mean survival time of MGP treated group was prolonged comparing with control group. Generally the group of intravenous injection is more effective than intraperitoneal injection. Conclusion : These results were suggested that MGP may be a useful anticancer agent for therapy of human lung cancer. And follow study need for the certain evidence.

천연형 사람 적혈구 조혈인자의 항원성시험 (Antigenicity Study of Recombinant Human Erythropoietin)

  • 강경구;조현;백남기;김원배
    • Biomolecules & Therapeutics
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    • 제6권1호
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    • pp.50-55
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    • 1998
  • Antigenic potential of a recombinant human erythropoietin (rhEPO) produced by Dong-A charm. Co. Ltd. was examined by active systemic anaphylaxis (ASA) test in guinea pigs, mouse-rat passive cutaneous anaphylaxis (PCA) reaction and passive hemagglutination (PHA) test. In ASA test, rhEPO induced the signs of restlessness, rubbing or licking nose, sneezing and coughing in the animals immunized with rhEPO 1000 lU/kg alone or rhEPO 1000 lU/kg incorporated into Freund\\\\`s complete adjuvant. In the mouse-rat PCA test, only one of six sera from the animals immunized with rhEPO 1000 lUng incorporated into Alum showed positive result. In the PHA test, rhEPO revealed negative results in all of the rhEPO-immunized groups. From these results, rhEPO was considered to produce IgE in guinea pigs and mice, but not IgG and/or IsM in mice. The results of this study were similar to those of the other recombinant human erythropoietin and these positive results were thought to be caused due to the fact that rhEPO were heterogeneous proteins to guinea pigs and mice. Considering the fact that rhErO has an identical structure with indigenous human erythropoietin, rhEPO is not thought to cause immunological problems in clinical use.

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DAZL binds to the transcripts of several Tssk genes in germ cells

  • Zeng, Mei;Deng, Wenqian;Wang, Xinying;Qiu, Weimin;Liu, Yanyan;Sun, Huaqin;Tao, Dachang;Zhang, Sizhong;Ma, Yongxin
    • BMB Reports
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    • 제41권4호
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    • pp.300-304
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    • 2008
  • The Dazl gene encodes a germ-cell-specific RNA-binding protein which is essential for spermatogenesis. It has been proposed that this protein (DAZL) binds to RNA in the cytoplasm of germ cells and controls spermatogenesis. Using the specific nucleic acids associated with proteins (SNAAP) technique, we identified 17 target mRNAs bound by mDAZL. Among these transcripts, we focused on TSSK2, which encodes a testis-specific serine/threonine kinase. To date, five TSSK family members have been cloned, and all are exclusively expressed in the testis. We demonstrated that in addition to the TSSK1 3'UTR, the 3'UTRs of TSSKs 2 and 4 were bound by human and mouse DAZL, and that human DAZL (hDAZL) bound to the 3'UTR of human TSSK5 (hTSSK5). Our results suggest that the Dazl gene may play different roles in human and mouse spermatogenesis by regulating different members of the downstream gene family.