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    Cell Fusion and Hybridization

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    Rachelle Cajes
    Cell fusion involves fusing two different cells into a single unit containing genes from both original cells. The most widely used fusion agent is Sendai virus, which is inactivated using UV light or B-propiolactone. To isolate pure populations of human-mouse hybrid cells, a selection procedure uses HAT medium to kill parental and homokaryon cells, allowing only hybrid cells containing human and mouse genes to survive and grow. Chromosomes are often lost preferentially after fusion, with up to 95% of human chromosomes lost in rodent-human fusions by the time hybrids are isolated.

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    Cell Fusion and Hybridization

    Uploaded by

    Rachelle Cajes
    72 views13 pages
    Cell fusion involves fusing two different cells into a single unit containing genes from both original cells. The most widely used fusion agent is Sendai virus, which is inactivated using UV light or B-propiolactone. To isolate pure populations of human-mouse hybrid cells, a selection procedure uses HAT medium to kill parental and homokaryon cells, allowing only hybrid cells containing human and mouse genes to survive and grow. Chromosomes are often lost preferentially after fusion, with up to 95% of human chromosomes lost in rodent-human fusions by the time hybrids are isolated.

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    Cell fusion involves fusing two different cells into a single unit containing genes from both original cells. The most widely used fusion agent is Sendai virus, which is inactivated using UV light or B-propiolactone. To isolate pure populations of human-mouse hybrid cells, a selection procedure uses HAT medium to kill parental and homokaryon cells, allowing only hybrid cells containing human and mouse genes to survive and grow. Chromosomes are often lost preferentially after fusion, with up to 95% of human chromosomes lost in rodent-human fusions by the time hybrids are isolated.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    72 views13 pages

    Cell Fusion and Hybridization

    Uploaded by

    Rachelle Cajes
    Cell fusion involves fusing two different cells into a single unit containing genes from both original cells. The most widely used fusion agent is Sendai virus, which is inactivated using UV light or B-propiolactone. To isolate pure populations of human-mouse hybrid cells, a selection procedure uses HAT medium to kill parental and homokaryon cells, allowing only hybrid cells containing human and mouse genes to survive and grow. Chromosomes are often lost preferentially after fusion, with up to 95% of human chromosomes lost in rodent-human fusions by the time hybrids are isolated.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 13

    Cell Fusion and

    Hybridization
    Reporter:
    Mapaa, Carrelle Joy T.
    Mamontayao, Jessievel
    Masugod, Aiza
    Cell Fusion
    Fusion of two different cells into a single unit
    which contains genes from both the original cell.

    The commonly used fusion agents for mapping of


    chromosome are Sendai virus, lysolecithin ,liposomes
    and polyethylene glycol (PEG).
    • The most widely used fusion agent is
    Sendai virus, which is inactivated by UV
    light or B-propiolactone.
    Selection of Hybrids
     To isolate pure population of Human-
    Mouse Heterokaryon a selection procedure
    is used that kills both the parental cells
    and the homokaryon but allows the human –
    mouse hybrid cells to survive and grow.
     In the medium contains a drug aminopterin
    , which blocks de novo purine and
    pyrimidine biosynthetic pathway of cells.
     The presence of hypoxanthine and
    thymidine the cells can overcome the
    block by synthezing and pyrimidines using
    salvage pathways.
     For cells to grow in HAT medium, two
    enzymes, hypoxanthine guanine
    phosphoribosyl transferase (HGPRT) and
    thymidine kinase (TK) must be functional.
    Heterokaryons , has normal HGRPT gene
    derived from the mouse genome and TK
    from gene from the human genome.
    Therefore, only hybrid cell grows.
    Chromosome Loss in Hybrid Cells
     When hybrid are formed between rodent
    and human cells, human chromosomes are
    lost preferentially.
     Loss of chromosome in interspecific
    hybrids was noted in fusion between mouse
    and rat cells , where chromosomes of rat
    were preferentially lost.
     In rodent and human cell fusions, up to
    95% of the human chromosomes can be
    lost by the time the hybrids are initially
    isolated and their chromosome analyzed.
    Stages of Cell Fusion:
    Stage I: Specification of the fusion fate
    Stage II: Cell attraction attachment and recognition
    Stage 3: Execution of Cell Membrane and
    Cytoplasmic Fusion
    Types of Cell Fusion
     Non Self Fusion – a cell fusion which allows
    the segregation of genes in new combinations
    into a single cell.
     Self-Fusion –
     Auto-Fusion
    Somatic Hybridization
    • Development of hybrid plants through the
    fusion of somatic protoplasts of two different
    plant species/varieties.
    Somatic Hybridization Technique
    • Isolation of Protoplasts
    • Fusion of the Protoplasts of desired
    species/varieties
    • Identification and Selection of somatic
    hybrid cells.
    • Culture of the hybrid cells.
    • Regeneration of Hybrid Plants
    Application of Somatic Hybridization
    • Opened new possibilities for the in vitro
    genetic manipulation of plants to improve the
    crops.
    • Helped in the successful transfer of some
    disease resistance genes.
    • Environmental Tolerance
    • Quality Character
    Problems and Limitations
    • Fertile and visible seeds are not always
    produced by plants .
    • The production of viable somatic hybrids is
    not possible in all instances even though
    Protoplasts fusion between different species
    or genus is easy.

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