PMID- 18423480
OWN - NLM
STAT- MEDLINE
DCOM- 20080709
LR  - 20210102
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 335
IP  - 1-2
DP  - 2008 Jun 1
TI  - Increased exosome production from tumour cell cultures using the Integra CELLine 
      Culture System.
PG  - 98-105
LID - 10.1016/j.jim.2008.03.001 [doi]
AB  - Exosomes are nanometer-sized vesicles, secreted from most cell types, with 
      documented immune-modulatory functions. Exosomes can be purified from cultured 
      cells but to do so effectively, requires maintenance of cells at high density in 
      order to obtain sufficient accumulation of exosomes in the culture medium, prior 
      to purification. Whilst high density cultures can be achieved with cells in 
      suspension, this remains difficult with adherent cells, resulting in low quantity 
      of exosomes for subsequent study. We have used the Integra CELLine culture 
      system, originally designed for hybridoma cultures, to achieve a significant 
      increase in obtainable exosomes from adherent and non-adherent tumour cells. 
      Traditional cultures of mesothelioma cells (cultured in 75 cm(2) flasks) gave an 
      average yield of 0.78 microg+/-0.14 microg exosome/ml of conditioned medium. The 
      CELLine Adhere 1000 (CLAD1000) flask, housing the same cell line, increased 
      exosome yield approximately 12 fold to 10.06 microg+/-0.97 microg/ml. The 
      morphology, phenotype and immune function of these exosomes were compared, and 
      found to be identical in all respects. Similarly an 8 fold increase in exosome 
      production was obtained from NKL cells (a suspension cell line) using a CELLine 
      1000 (CL1000) flask. The CELLine system also incurred ~5.5 fold less cost and 
      reduced labour for cell maintenance. This simple culture system is a cost 
      effective, useful method for significantly increasing the quantity of exosomes 
      available from cultured cells, without detrimental effects. This tool should 
      prove advantageous in future studies of exosome-immune modulation in cancer and 
      other settings.
FAU - Mitchell, J Paul
AU  - Mitchell JP
AD  - Department of Oncology & Palliative Medicine, School of Medicine, Cardiff 
      University, Velindre Cancer Centre, Whitchurch, Cardiff, UK. 
      paul.mitchell@velindre-tr.wales.nhs.uk
FAU - Court, Jacqueline
AU  - Court J
FAU - Mason, Malcolm David
AU  - Mason MD
FAU - Tabi, Zsuzsanna
AU  - Tabi Z
FAU - Clayton, Aled
AU  - Clayton A
LA  - eng
PT  - Journal Article
DEP - 20080403
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Culture Media, Conditioned)
SB  - IM
MH  - Cell Adhesion
MH  - *Cell Culture Techniques
MH  - Cell Line
MH  - Cell Proliferation
MH  - Culture Media, Conditioned/metabolism
MH  - Humans
MH  - Killer Cells, Natural/immunology/*metabolism/pathology
MH  - Leukemia, Large Granular Lymphocytic/immunology/*metabolism/pathology
MH  - Mesothelioma/immunology/*metabolism/pathology
MH  - Microscopy, Electron, Transmission
MH  - Phenotype
MH  - Pleural Neoplasms/immunology/*metabolism/pathology
MH  - Secretory Vesicles/immunology/*metabolism/ultrastructure
MH  - Time Factors
MH  - Tumor Cells, Cultured
EDAT- 2008/04/22 09:00
MHDA- 2008/07/10 09:00
CRDT- 2008/04/22 09:00
PHST- 2007/11/06 00:00 [received]
PHST- 2008/03/03 00:00 [revised]
PHST- 2008/03/05 00:00 [accepted]
PHST- 2008/04/22 09:00 [pubmed]
PHST- 2008/07/10 09:00 [medline]
PHST- 2008/04/22 09:00 [entrez]
AID - S0022-1759(08)00092-6 [pii]
AID - 10.1016/j.jim.2008.03.001 [doi]
PST - ppublish
SO  - J Immunol Methods. 2008 Jun 1;335(1-2):98-105. doi: 10.1016/j.jim.2008.03.001. 
      Epub 2008 Apr 3.