Animal cells reversibly permeable to small molecules

Area of Science:

• Cell Biology
• Biochemistry
• Metabolism Regulation

Background:

• Studying cellular metabolism requires methods to introduce substrates into cells.
• Existing methods often compromise cell integrity or fail to penetrate essential cellular components.

Purpose of the Study:

• To develop a novel cell preparation technique for studying metabolic regulation.
• To characterize the properties and utility of these permeable cells in biochemical assays.

Main Methods:

• Baby hamster kidney cells were made permeable via hypertonic treatment.
• The permeable cells' ability to synthesize DNA, RNA, and protein was assessed.
• Reversible permeability and resealing capabilities were investigated.
• The effect of hydroxyurea on DNA synthesis was examined in permeable cells.

Main Results:

• Permeable cells retained morphology, organelles, DNA, and most protein.
• Synthesized DNA, RNA, and protein at rates comparable to intact cells.
• Demonstrated cell-cycle dependent DNA replication and active ribonucleotide reductase.
• Hydroxyurea's mechanism as a ribonucleotide reductase inhibitor was confirmed.
• Reversible permeability allowed incorporation of compounds, demonstrating potential for drug studies.

Conclusions:

• The developed permeable cell preparation is a valuable tool for metabolic studies.
• This method allows investigation of poorly penetrating compounds, including drugs.
• The reversible nature of permeability enhances its utility in various cellular research applications.