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第三章 动物细胞工程制药 PowerPoint PPT Presentation


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第三章 动物细胞工程制药. 一切生命的关键问题都要到细胞中去寻找. 第一节 概 述. 1665 年,英国物理学家 Robert Hooke 用自制的显微镜首次发现了细胞,奠定了细胞生物学的基础 。. 荷兰科学家 Leeuwen hooke 用设计较好的显微镜观察了许多动植物的活细胞与原生动物. 细胞学说创立. 动物细胞与组织培养的发展历史. 组织和细胞培养的早期萌芽阶段 1885 年,德国人 Roux 用生理盐水培养鸡胚组织,使之存活了数月之久 1897 年, Loeb 证明从血液和结缔组织中分离到的细胞可在血清和血浆中存活

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第三章 动物细胞工程制药

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1665Robert Hooke


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Leeuwen hooke


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    • 1885Roux

    • 1897Loeb

    • 1903Jolly


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    • 1907Harrison

    • Carrel1923


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sheep

Sheep-goat chimera

goat


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(tPA) CHO 1987

(EPO) CHO 1989

(hGF) C127 1989

(G-CSF) CHO 1991

III CHO A 1992-1993

CHO 1996

CHO 1994

CHO 1998

CHOChinese hamster ovary

C127: Mouse fibroblast


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  • cell fusion


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A. Bread wheat ; B. Rye

C. Wheat and rye are crossed together to produce the hybrid triticale.

The massive taproot of wild radish ,a common naturalized weed in San Diego County, California. Unlike the numerous, tender, cultivated varieties of radishes, wild radish has a tough, woody taproot that is unpalatable


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Scientists change the genes in living cells by putting the desired new gene into a little virus-like organism which is allowed to get into your cells and which inserts the new gene into the cell along with the old genes.


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Manipulation of a cattle embryo and twin calves

Test- tube baby


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  • DNARNA


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DNA


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DNA


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G1DNA

SDNA

G2DNA

M

Tc G1 + S + G2 + M


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  • anchorage-dependent

  • anchorage-independent


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Fibroblast like cell type

fibroblast like cell type

  • 23


Epithilium like cell type

epithilium-like cell type


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Hela

pH pH

3T3


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CHO

L929


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  • 1248

  • pH


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  • ( contact inhibition


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  • density inhibition


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Hayflick


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  • pH


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  • 128


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Life Span of Culture Cells


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  • 14


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  • cell line


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  • 2ndiploid cell line

  • 50


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MRC-5

WI-38


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Transformation

transformation


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  • SV40


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293T human embryonal kidney, SV40 and large T antigen transformed

Veromonkey kidney

BHK-21Kidney Fibroblast Cells

CHO-K1


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  • passage number

  • generation number


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  • pH


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pH


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CO2


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  • MEMDMEMIMDM RPMI1640

    199109HamF12


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  • BBSHanksEarle

  • pH NaHCO3HEPES

  • EDTA

  • -


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  • suspension culture

  • anchorage-dependent culture


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  • microcarrier culture

  • porous microcarrier


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  • batch

  • fed-batch

  • semi-continuous

  • continuous

  • perfusion


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  • 107109/ml


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