• 제목/요약/키워드: stable cells

검색결과 968건 처리시간 0.031초

High-Level Expression of Recombinant Human Bone Morphogenetic Protein-4 in Chinese Hamster Ovary Cells

  • PARK JUNHO;YU SUNGRYUL;YOON JAESEUNG;BAEK KWANGHEE
    • Journal of Microbiology and Biotechnology
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    • 제15권6호
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    • pp.1397-1401
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    • 2005
  • Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

Defective Interfering HIV-1 Pseudotypes Carrying Chimeric CD4 Protein

  • Park, Seung-Won;Ye, Zhiping;Schubert, Manfred;Paik, Soon-Young
    • BMB Reports
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    • 제34권6호
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    • pp.566-572
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    • 2001
  • Chimeric CD4 proteins were assembled. They contained the entire CD4 ectodomain that is linked to different membrane anchors. Membrane anchors consisted of either glucosyl phosphatidyl inositol (gpi), the transmembrane and cytoplasmic regions of HIV-1 Env protein, or the vesicular stomatitis virus G glycoprotein, respectively. The HIV-1 co-receptor CXCR4 and CD4 were independently inserted into viral envelopes. We compared the insertion of six different CD4/CXCR4 constructs into HIV-1 envelopes, as well as their functionality in targeting and specific infection of cells that constitutively express the HIV-1 Env protein. All of the six different HIV-1 (CD4/CXCR4) pseudotypes were able to transduce Env (+) cells at similar efficiency. In addition, stable transduction of the Env (+) recipient cells demonstrated that all chimeric proteins were functional as receptors for Env when inserted into HIV-1 envelopes. In fact, these results demonstrate for the first time a stable transduction by a targeted HIV-1 pseudotype virus.

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GLOBAL STABILITY OF HIV INFECTION MODELS WITH INTRACELLULAR DELAYS

  • Elaiw, Ahmed;Hassanien, Ismail;Azoz, Shimaa
    • 대한수학회지
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    • 제49권4호
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    • pp.779-794
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    • 2012
  • In this paper, we study the global stability of two mathematical models for human immunodeficiency virus (HIV) infection with intra-cellular delays. The first model is a 5-dimensional nonlinear delay ODEs that describes the interaction of the HIV with two classes of target cells, $CD4^+$ T cells and macrophages taking into account the saturation infection rate. The second model generalizes the first one by assuming that the infection rate is given by Beddington-DeAngelis functional response. Two time delays are used to describe the time periods between viral entry the two classes of target cells and the production of new virus particles. Lyapunov functionals are constructed and LaSalle-type theorem for delay differential equation is used to establish the global asymptotic stability of the uninfected and infected steady states of the HIV infection models. We have proven that if the basic reproduction number $R_0$ is less than unity, then the uninfected steady state is globally asymptotically stable, and if the infected steady state exists, then it is globally asymptotically stable for all time delays.

Expression and Characterization of Purinergic Receptor, $P2Y_{10}$ in Hematopoietic Stem Cells

  • Lee Eun-Jong;Kim Dong-Ku
    • Reproductive and Developmental Biology
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    • 제29권2호
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    • pp.109-115
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    • 2005
  • Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughoutthe lifetime. In this study, we analyzed the expression of receptors of $P2Y_{10}$, purinergic receptor families in murine hematopoietic stem cells, hematopoietic progenitor cells. In addition, the biological activity of $P2Y_{10}$ was investigated with B lymphocyte cell line, Ba/F3 in effect to cell growth and cell cycle. From the analysis of expression in hematopoieticstem cell. and progenitor with RT-PCR, $P2Y_{10}$ was strongly expressed in murine hematopoieticstem cells (c-kit+ Sca-l+ Lin-) and progenitor cell population, such as c-kit- Sca-l+ Lin-, c-kit+ Sca-l- Lin- and c-kit- Sca-l- Lin-. To investigate the biological effects by $P2Y_{10}$, retroviral vector from subcloned murine $P2Y_{10}$ cDNA was used fur gene introduction into Ba/F3 cells, and stable transfectant cells were obtained by flow cytometry sorting. In cell proliferation assay, the proliferation ability of $P2Y_{10}$ receptor gene­transfected cells was strongly inhibited, and the cell cycle was arrested at G1 phase. These result suggest that the $P2Y_{10}$ may be involved the biological activity in hematopoietic stem cells and immature B lymphocytes.

배아추출물 공여 배아의 발생단계가 바다송사리(Oryzias dancena) 배아 줄기세포 유사세포의 배양에 미치는 영향 (Effects of the Developmental Stage of Extract Donor Embryos on the Culture of Marine Medaka Oryzias dancena Embryonic Stem Cell-like Cells)

  • 류준형;공승표
    • 한국수산과학회지
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    • 제50권2호
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    • pp.160-168
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    • 2017
  • Optimizing the conditions for stem cell culture is an essential prerequisite for the efficient utilization of stem cells. In the culture of fish embryonic stem cells (ESCs) or ESC-like cells, embryo extracts are important for stable growth, but there is no rule for determining the developmental stage of the embryos used to obtain extracts. Therefore, this study investigated the effects of the developmental stage of extract donor embryos on the culture of Oryzias dancena ESC-like cells. O. dancena ESC-like cells were cultured in different media containing each of four types of embryo extract depending on the developmental stage of the extract donor embryos. Growth, morphology, colony-forming ability, alkaline phosphatase (AP) activity, and embryoid body (EB) formation of the cells were investigated. While the developmental stage of the extract donor embryos did not influence the growth, morphology, AP activity, or EB formation of ESC-like cells, colony-forming ability was affected and the pattern of the effects differed completely between the two ESC-like cells investigated. These results suggest that the developmental stage of extract donor embryos should be selected carefully for the culture of ESC-like cells, according to the research purpose and type of cell line.

항체를 포함하는 Phosphatidylethanolamine 리포좀의 제조와 그 특성 (Preparation and Characterization of PE Liposomes Containing Antibody)

  • 박성호;신현재양지원최태부
    • KSBB Journal
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    • 제10권2호
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    • pp.204-211
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    • 1995
  • 인지질 그 자체만으로는 안정한 이중층 리포좀을 형성하지 못하는 불포화 PE(DOPE)에 palmitoyl가 가 결합된 항체(p-IgG)를 지질층에 삽입시켜 lmm t unoliposome을 제조하고 그 특성에 관하여 살펴 보 았다. 우선 안정된 리포좀을 제조하기 위해서 고려 해야할 인자들로 항체 가공방법, 지질과 항체와의 몰 비, 그리고 각종 첨가제들에 대한 최적 조건을 조 사하였다. 예를 들면 p-IgG와 lipid의 볼비를 $2.5{\times}10^{-4}$ 으로 했을 때 안정한 리포좀을 만들 수 있었으며, 첨가제로 들어가는 DOC의 경우 최종 농도가 O.09wt % 일 때 calcein의 포집률이 최대가 되었고 c calcein의 최종 pH는 8.5~9.5 정도에서 안정한 라포좀이 제조될 수 있었다. 다중 빛 단일클론의 항체 를 삽입한 리포좀을 표면항원을 가진 표적세포와 결합시켰을 때 리포좀이 와해되면셔 포집된 calcein이 방출되는 것으로 보아 삽입된 p-IgG가 PE 리포좀을 형성하는데 필수척임을 알 수 있었다. 또 같은 리포좀을 비특이적인 세포와 접촉시켰을 때에는 아무 런 변화를 보이지 않아 calcein 방출이 항원-항체 반응에 의한 것임을 알 수 있었고 이로부터 표적 민 감성 PE 리포좀이 만들어졌음을 확인하였다.

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머신러닝 기반 스마트 단말기 Lithium-Ion Cell의 잔량 추정 방법의 실증적 연구 (An Empirical Study on Machine Learning based Smart Device Lithium-Ion Cells Capacity Estimation)

  • 장성진
    • 문화기술의 융합
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    • 제6권4호
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    • pp.797-802
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    • 2020
  • 지난 몇 년 동안 스마트 폰을 비롯한 다양한 스마트 기기들은 휴대성을 기반으로 사용자의 요구에 의해 지속적으로 성능이 향상 되고 있다. 유비쿼터스 컴퓨팅 (Ubiquitous Computing) 환경과 센서 네트워크 (Sensor network)등의 다양한 망 접속 기술로 인하여 휴대성을 기반으로 하는 단말기들이 다양하게 보급되어 사용되고 있다. 스마트 단말들은 사용 중에 보다 안정적인 동작을 위하여 에너지 모니터링을 세밀하게 할 수 있는 기술이 필요하게 되었다. 소형 경량화 된 스마트 단말기는 다양한 멀티미디어 작업으로 인하여 단말 운용 중에 전원 부족 문제가 발생하게 된다. 이와 같은 상황을 미리 방지하고 안정적인 단말 운용을 위해서 기존에 다양한 추정 하드웨어가 개발 되었다. 그러나 잔량 추정을 하는 방법이나 성능이 비교적 우수하지 못하였다. 본 논문에서는 스마트 단말의 운용 중에 발생 할 수 있는 잔여 잔량 문제를 미리 예측하여 보다 안정적인 운용을 위한 리튬이온 셀의 잔량 추정 방법을 머신러닝에 기초를 두고 연구 하였다. 기존의 하드웨어적인 추정 방법이 아니라 사용 중인 리튬이온 셀의 특성을 머신러닝 기법을 이용한 학습 알고리즘으로 학습 시키고 최적화된 결과를 추정하여 적용 하고자 한다.

Production of Acrylamide Using Immobilized Cells of Rhodococcus rhodochrous M33

  • Kim, Bu-Youn;Hyun, Hyung-Hwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권4호
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    • pp.194-200
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    • 2002
  • The cells of Rhodococcus rhodochrous M33, which produce a nitrile hydratase enzyme, were immobilized in acrylamide-based polymer gels. The optimum pH and temperature for the activity of nitrile hydratase in both the free and Immobilized cells were 7.4 and 45$\^{C}$, respectively, yet the optimum temperature for acrylamide production by the immobilized cells was 20$\^{C}$. The nitrile hydratase of the immobilized cells was more stable with acrylamide than that of the free cells. Under optimal conditions, the final acrylamide concentration reached about 400 g/L with a conversion yield of almost 100% after 8 h of reaction when using 150 g/L of immobilized cells corresponding to a 1.91 g-dry cell weight/L. The enzyme activity of the immobilized cells rapidly de-creased with repeated use. However, the quality of the acrylamide produced by the immobilized cells was much better than that produced by the free cells in terms of color, salt content, turbidity, and foam formation. The quality of the aqueous acrylamide solution obtained was found to be of commercial use without further purification.

Establishment of An Efficient and Stable Transgene Expression System in Chicken Primordial Germ Cells

  • Yang, Ju-Hyun;Kim, Sung-Tae
    • Bulletin of the Korean Chemical Society
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    • 제33권5호
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    • pp.1536-1540
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    • 2012
  • Chicken primordial germ cells (cPGCs) are founder germ cells in embryonic stage of development that eventually give rise to sperms or oocytes. Currently cPGCs are only known cells enabling germline transmission in chicken and their cultivation protocols were recently established. Although genome modifications of chickens are now theoretically possible using cPGCs, there are still several hurdles to overcome to practically use cPGCs as mediators for chicken transgenesis. First, efficiency of gene delivery into cPGCs remains low with current methods. Second, there aregene silencing mechanisms against the expression of foreign genes in cPGCs. In this study, we successfully increased the efficiency of gene delivery in cPGCs by taking advantage of the TTAA-specific $piggybac$ transposon system. Moreover, a pipette-type electroporator significantly enhanced transfection efficiency up to 5-fold compared withcuvette-type methods. Taken together, the technological advances in our study will provide practical benefits for the application to fulfill genetic modifications of chicken genome.