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Cell-cell Fusion of Genome Edited Cell Lines for Perturbation of Cellular Structure and Function
Published on: December 7, 2019
Chromosome transfer via cell fusion.
1Istituto di Ricerca Genetica e Biomedica, Consiglio Nazionale delle Ricerche, Monserrato, Italy. marianna.paulis@humanitas.it
Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
Summary
Microcell-mediated chromosome transfer (MMCT) and micronucleated whole cell fusion (MWCF) enable chromosome transfer between cells. MWCF offers higher efficiency and simplicity compared to MMCT, but is limited to interspecies applications.
Area of Science:
- Cell biology
- Genetics
- Molecular biology
Background:
- Chromosome transfer into recipient cells is crucial for various biological applications.
- Microcell-mediated chromosome transfer (MMCT) is a standard method but suffers from low efficiency.
- Alternative methods are needed to improve chromosome transfer efficiency and simplicity.
Purpose of the Study:
- To describe and compare two chromosome transfer protocols: MMCT and micronucleated whole cell fusion (MWCF).
- To highlight the advantages and limitations of each method for chromosome delivery into cells.
- To provide insights into optimizing chromosome transfer efficiency based on parental cell line selection.
Main Methods:
- Detailed description of the microcell-mediated chromosome transfer (MMCT) protocol.
- Detailed description of the micronucleated whole cell fusion (MWCF) protocol, a novel approach without microcell isolation.
- Evaluation of chromosome transfer efficiency based on the choice of donor and recipient cell lines.
Main Results:
- MWCF demonstrates significantly higher chromosome transfer efficiency compared to MMCT.
- MWCF is simpler to perform than MMCT due to the absence of microcell isolation.
- MMCT can be adapted for any donor and recipient cell line, while MWCF is limited to interspecies applications.
Conclusions:
- MWCF represents a more efficient and simpler alternative to MMCT for chromosome transfer, particularly for interspecies applications.
- The efficiency of both MMCT and MWCF is highly dependent on the specific parental cell lines used.
- Both methods facilitate the transfer of intact chromosomes or fragments into recipient cells without affecting their viability or chromosomal stability.
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