Efficient and reproducible generation of high-expressing, stable human cell lines without need for antibiotic selection

Schiedner, Gudrun; Hertel, Sabine; Bialek, Corinna; Kewes, Helmut; Waschütza, Gero; Volpers, Christoph

doi:10.1186/1472-6750-8-13

Cited by 43 publications

(24 citation statements)

References 40 publications

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“…We started with a population of the HEK-293 cells transfected by a GFP-encoding plasmid that could not replicate inside the cells and contained no sequences promoting its integration into the chromosomes. Furthermore, we did not use antibiotic or any other form of chemical selection that could affect the cell physiology or result in cell line instability (34,35) nor used a cell sorter to enrich the population by GFP-producing cells.…”

Section: Resultsmentioning

confidence: 99%

Planar Arrangement of Eukaryotic Cells in Merged Hydrogels Combines the Advantages of 3-D and 2-D Cultures

2012

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We report an unordered 2-D array of eukaryotic cells completely embedded in a 3-D matrix. Every cell is located at the same distance from the gel surface, which ensures uniformity of growth conditions and ease of observation characteristic of a 2-D culture. Each cell is firmly immobilized, and each has a unique address in the array. The cells can be rapidly screened, individually monitored during extended time periods, and cultured with the formation of spheroid microcolonies characteristic of a 3-D culture. Individual microcolonies can be extracted from the gel and further propagated, thus enabling isolation of pure cell clones from rather dense cell populations and rapid drug-free generation of stable cell lines.

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Section: Resultsmentioning

confidence: 99%

Planar Arrangement of Eukaryotic Cells in Merged Hydrogels Combines the Advantages of 3-D and 2-D Cultures

2012

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“…Upon immortalization with the abovementioned adenoviral genes E1A/E1B and pIX a cell line was established designated as CAP [120,121]. Although the specific cell type that Brought to you by | New York University Bobst Library Technical Services Authenticated Download Date | 7/6/15 11:50 AM is immortalized by this procedure has not yet been defined, these cells have been shown to give rise to high levels of protein [121,122] as well as influenza virus [123].…”

Section: Recent Developmentsmentioning

confidence: 99%

2.1 Generation of Cell Lines and Biotechnological Applications

Lipps¹,

May²,

Wirth³

2014

Animal Cell Biotechnology

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“…To extend the replicative life span, human amniocytes have been immortalised by adenovirus E1A-, E1B-and pIX-functions. The cells were primarily developed for the stable production of recombinant proteins and it was proven that complex proteins can be expressed carrying a high degree of human glycosylation patterns [32].…”

Section: Capmentioning

confidence: 99%