PMID- 22359005
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20121002
LR  - 20211021
IS  - 0920-9069 (Print)
IS  - 1573-0778 (Electronic)
IS  - 0920-9069 (Linking)
VI  - 26
IP  - 1
DP  - 1998 Jan
TI  - Calcium-phosphate mediated DNA transfer into HEK-293 cells in suspension: control 
      of physicochemical parameters allows transfection in stirred media. Transfection 
      and protein expression in mammalian cells.
PG  - 39-47
LID - 10.1023/A:1007917318181 [doi]
AB  - This is the first report of transient transfection of suspended cells with 
      purified plasmid DNA in bioreactors or spinner flasks. DNA/calcium phosphate 
      complexes were pumped or injected directly into stirred cultures of the 
      immortalized human embryo kidney cell line 293 (HEK-293) which had been adapted 
      to growth in suspension. We identified culture conditions suitable for this 
      approach and modified the protocol for the generation of precipitate complexes, 
      based on our earlier work. In order to stabilize the 'DNA-vehicle'-complex in the 
      culture medium, we identified pH ranges and ion-concentrations which prevent 
      dissolution or aggregation of the precipitate particles. Such conditions 
      maintained suspended fine particles in spinners or bioreactors for up to 6 hr. 
      During that period, cells and precipitate complexes interacted sufficiently to 
      allow DNA transfer and subsequent expression of recombinant protein. In a simple 
      5 day batch process, with a starting density of 0.3 x 10(6) cells mL(-1), about 
      0.5 mg L(-1) of a recombinant tissue plasminogen activator variant was observed.
FAU - Jordan, M
AU  - Jordan M
AD  - Dept. Chemistry, Swiss Federal Institute of Technology Lausanne, CH-1015, 
      Lausanne, Switzerland.
FAU - Kohne, C
AU  - Kohne C
FAU - Wurm, F M
AU  - Wurm FM
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Cytotechnology
JT  - Cytotechnology
JID - 8807027
PMC - PMC3449510
EDAT- 1998/01/01 00:00
MHDA- 1998/01/01 00:01
PMCR- 1999/01/01
CRDT- 2012/02/24 06:00
PHST- 2012/02/24 06:00 [entrez]
PHST- 1998/01/01 00:00 [pubmed]
PHST- 1998/01/01 00:01 [medline]
PHST- 1999/01/01 00:00 [pmc-release]
AID - 141410 [pii]
AID - 10.1023/A:1007917318181 [doi]
PST - ppublish
SO  - Cytotechnology. 1998 Jan;26(1):39-47. doi: 10.1023/A:1007917318181.