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Related Concept Videos

Cell Lines01:16

Cell Lines

A cell line is a population of cells grown in vitro that can be subcultured over several generations. Normal cells cease to divide after a certain number of cell divisions, a process known as replicative senescence. This number, called the Hayflick limit, was conceptualized by Leonard Hayflick in 1961 when he observed that fetal cells grown in culture could only divide 40-60 times. This limit is due to the shortening of the telomeres during each round of cell division, preventing cell division...
Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

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Related Experiment Video

Updated: Jun 2, 2026

A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics
07:48

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Published on: September 22, 2011

Methods to create a stringent selection system for mammalian cell lines.

H J M Van Blokland1, F Hoeksema, M Siep

  • 1Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH, Amsterdam, The Netherlands.

Cytotechnology
|April 22, 2011
PubMed
Summary

This study introduces a stringent selection system for high-protein producing mammalian cell lines using the Zeocin resistance marker. Modified Zeocin systems enable faster development of cell lines with superior protein expression levels.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Efficient generation of high-protein producing recombinant mammalian cell lines is crucial for biopharmaceutical production.
  • Stringent selection systems are essential for isolating cell lines with optimal protein expression.

Purpose of the Study:

  • To develop a more stringent selection system for mammalian cell lines using the Zeocin resistance marker.
  • To improve the efficiency and speed of establishing high-protein producing cell lines.

Main Methods:

  • Cloning DNA sequences of varying lengths upstream of the Zeocin resistance gene to modulate translation efficiency.
  • Performing genetic screens to identify mutations in the Zeocin resistance protein that impair its function.
  • Combining modified Zeocin systems with short peptides to enhance selection stringency.

Main Results:

  • Increasing the length of upstream translated peptides decreased Zeocin marker expression, increasing selection stringency.
  • Identified Zeocin mutants that functionally impair the selection marker.
  • Combining longer peptides or Zeocin mutants with short peptides resulted in fewer, but higher-protein producing, transfected colonies.

Conclusions:

  • The developed Zeocin-based selection system offers enhanced stringency for mammalian cell line development.
  • This system facilitates the rapid establishment of high-yield recombinant protein-producing cell lines.
  • The flexible system allows for precise control over selection stringency and protein expression levels.