A Serum-Free, Transient Transfection System for Enhancing Production of Recombinant Antibodies in Mammalian Cells

2010 ◽  
pp. 229-232
Author(s):  
Gaurav Backliwal ◽  
Sarah Wulhfard ◽  
Fanny Delegrange ◽  
Lucia Baldi ◽  
Maria de Jesus ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Recombinant Antibodies ◽  
Transient Transfection ◽  
Serum Free ◽  
Enhancing Production

Serum-free large-scale transient transfection of CHO cells

2004 ◽  
Vol 87 (4) ◽  
pp. 537-545 ◽  
Author(s):  
Madiha Derouazi ◽  
Philippe Girard ◽  
Fr�d�ric Van Tilborgh ◽  
Keyvan Iglesias ◽  
Natalie Muller ◽  
...  
Keyword(s):  
Cho Cells ◽  
Large Scale ◽  
Transient Transfection ◽  
Serum Free

scholarly journals Specific Double-Stranded RNA Interference in Undifferentiated Mouse Embryonic Stem Cells

2001 ◽  
Vol 21 (22) ◽  
pp. 7807-7816 ◽  
Author(s):  
Shicheng Yang ◽  
Stephen Tutton ◽  
Eric Pierce ◽  
Kyonggeun Yoon
Keyword(s):  
Gene Expression ◽  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Es Cells ◽  
Embryonic Stem ◽  
Transient Transfection ◽  
Interferon Response ◽  
Green Fluorescent Protein Gene ◽  
Double Stranded Rna ◽  
Rnai Activity

ABSTRACT Specific mRNA degradation mediated by double-stranded RNA (dsRNA) interference (RNAi) is a powerful way of suppressing gene expression in plants, nematodes, and fungal, insect, and protozoan systems. However, only a few cases of RNAi have been reported in mammalian systems. Here, we investigated the feasibility of the RNAi strategy in several mammalian cells by using the enhanced green fluorescent protein gene as a target, either by in situ production of dsRNA from transient transfection of a plasmid harboring a 547-bp inverted repeat or by direct transfection of dsRNA made by in vitro transcription. Several mammalian cells including differentiated embryonic stem (ES) cells did not exhibit specific RNAi in transient transfection. This long dsRNA, however, was capable of inducing a sequence-specific RNAi for the episomal and chromosomal target gene in undifferentiated ES cells. dsRNA at 8.3 nM decreased the cognate gene expression up to 70%. However, RNAi activity was not permanent because it was more pronounced in early time points and diminished 5 days after transfection. Thus, undifferentiated ES cells may lack the interferon response, similar to mouse embryos and oocytes. Regardless of their apparent RNAi activity, however, cytoplasmic extracts from mammalian cells produced a small RNA of 21 to 22 nucleotides from the long dsRNA. Our results suggest that mammalian cells may possess RNAi activity but nonspecific activation of the interferon response by longer dsRNA may mask the specific RNAi. The findings offer an opportunity to use dsRNA for inhibition of gene expression in ES cells to study differentiation.

scholarly journals Establishment of fast-growing serum-free immortalised cells from Chinese hamster lung tissues for biopharmaceutical production

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Noriko Yamano-Adachi ◽  
Rintaro Arishima ◽  
Sukwattananipaat Puriwat ◽  
Takeshi Omasa
Keyword(s):  
Cell Line ◽  
Cho Cells ◽  
Mammalian Cells ◽  
High Efficiency ◽  
Chinese Hamster ◽  
Serum Free ◽  
Sterility Testing ◽  
Fast Growing ◽  
Defined Culture

Abstract Chinese hamster (Cricetulus griseus) ovary-derived Chinese hamster ovary (CHO) cells are the most commonly used mammalian hosts for the industrial production of recombinant therapeutics because of their ability to fold, assemble, and perform post-translational modifications, such as glycosylation, on proteins. They are also valuable for their ability to grow in serum-free suspension cultures. In this study, we established a cell line derived from lung tissue of Chinese hamsters, named Chinese hamster lung (CHL)-YN cells. The biosafety of CHL-YN cells was confirmed by in vitro sterility testing, mycoplasma detection, and reverse transcriptase assays. One of the key characteristics of CHL-YN cells was their doubling time of 8.1 h in chemically defined culture medium; thus, they proliferate much faster than conventional CHO cells and general mammalian cells. Transgenes could be introduced into CHL-YN cells with high efficiency. Finally, between 50% to > 100% of the amount of glycosylated immunoglobulin G (IgG)1 produced by CHO-K1 cells was produced by CHL-YN cells over a shorter period of time. In summary, fast-growing CHL-YN cells are a unique cell line for producing recombinant proteins.

Transient Transfection of Mammalian Cells

2019 ◽  
pp. 143-153
Author(s):  
Susan Carson ◽  
Heather B. Miller ◽  
D. Scott Witherow ◽  
Melissa C. Srougi
Keyword(s):  
Mammalian Cells ◽  
Transient Transfection

Large-scale transient transfection of serum-free suspension-growing HEK293 EBNA1 cells: Peptone additives improve cell growth and transfection efficiency

2003 ◽  
Vol 84 (3) ◽  
pp. 332-342 ◽  
Author(s):  
Phuong Lan Pham ◽  
Sylvie Perret ◽  
Huyen Chau Doan ◽  
Brian Cass ◽  
Gilles St-Laurent ◽  
...  
Keyword(s):  
Cell Growth ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Transient Transfection ◽  
Serum Free ◽  
Serum Free Suspension

Receptor dependent cellular uptake of synthetic low density lipoprotein by mammalian cells in serum-free tissue culture

2006 ◽  
Vol 58 (10) ◽  
pp. 1337-1342 ◽  
Author(s):  
Sima Hayavi ◽  
George Baillie ◽  
Moira D. Owens ◽  
Gavin W. Halbert
Keyword(s):  
Tissue Culture ◽  
Cellular Uptake ◽  
Mammalian Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Serum Free

Serum-free growth medium for the cultivation of a wide spectrum of mammalian cells in stirred bioreactors

1988 ◽  
Vol 1 (4) ◽  
pp. 319-329 ◽  
Author(s):  
Volker J�ger ◽  
J�rgen Lehmann ◽  
Peter Friedl
Keyword(s):  
Growth Medium ◽  
Mammalian Cells ◽  
Wide Spectrum ◽  
Serum Free ◽  
Free Growth ◽  
Stirred Bioreactors

Rakkyo fructan as a cryoprotectant for serum-free cryopreservation of mammalian cells

2014 ◽  
Vol 118 (1) ◽  
pp. 101-106 ◽  
Author(s):  
Akiko Ogawa ◽  
Shinya Mizui ◽  
Yasuhito Chida ◽  
Masafumi Shimizu ◽  
Satoshi Terada ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Serum Free
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