Cell fusion-impaired variants of a mouse lymphocytic cell line obtained by single-step selection for polyethylene...

Area of Science:

• Cell biology
• Genetics
• Biochemistry

Background:

• Intercellular plasma membrane fusion is crucial for various biological processes.
• Understanding the genetic and molecular basis of fusion regulation is important.
• Previous methods for isolating fusion-impaired variants were less efficient.

Purpose of the Study:

• To develop an effective single-step method for isolating cell line variants with reduced intercellular plasma membrane fusion ability.
• To characterize the phenotype of these variants, including their stability and fusion competence.

Main Methods:

• Selection of L5178Y cell sublines for growth in toxic levels of polyethylene glycol (PEG) 6000.
• Induction of variants through ethyl methanesulfonate (EMS) treatment and spontaneous occurrence.
• Assessment of fusion competence and stability of isolated variants through serial subculture.

Main Results:

• Successfully isolated cell variants with significantly reduced intercellular fusion ability.
• Identified a pleiotropic phenotype including fusion impairment and PEG resistance (PgfR).
• Demonstrated stable PgfR phenotype retention for over 165 days in nonselective medium.
• Quantified an approximate 7-fold reduction in intercellular fusion competence in PgfR variants.

Conclusions:

• The described method provides an effective single-step approach for isolating cell variants with impaired fusion.
• The PgfR phenotype is stable and characterized by reduced intercellular fusion competence and PEG resistance.
• These variants serve as valuable tools for studying the mechanisms regulating cell fusion.