PMID- 20003054
OWN - NLM
STAT- MEDLINE
DCOM- 20100505
LR  - 20171116
IS  - 1537-2995 (Electronic)
IS  - 0041-1132 (Linking)
VI  - 50
IP  - 4
DP  - 2010 Apr
TI  - Evaluating maturation and genetic modification of human dendritic cells in a new 
      polyolefin cell culture bag system.
PG  - 843-55
LID - 10.1111/j.1537-2995.2009.02520.x [doi]
AB  - BACKGROUND: Dendritic cells (DCs) are applied worldwide in several clinical 
      studies of immune therapy of malignancies, autoimmune diseases, and 
      transplantations. Most legislative bodies are demanding high standards for 
      cultivation and transduction of cells. Closed-cell cultivating systems like cell 
      culture bags would simplify and greatly improve the ability to reach these 
      cultivation standards. We investigated if a new polyolefin cell culture bag 
      enables maturation and adenoviral modification of human DCs in a closed system 
      and compare the results with standard polystyrene flasks. STUDY DESIGN AND 
      METHODS: Mononuclear cells were isolated from HLA-A*0201-positive blood donors by 
      leukapheresis. A commercially available separation system (CliniMACS, Miltenyi 
      Biotec) was used to isolate monocytes by positive selection using CD14-specific 
      immunomagnetic beads. The essentially homogenous starting cell population was 
      cultivated in the presence of granulocyte-macrophage-colony-stimulating factor 
      and interleukin-4 in a closed-bag system in parallel to the standard flask 
      cultivation system. Genetic modification was performed on Day 4. After induction 
      of maturation on Day 5, mature DCs could be harvested and cryopreserved on Day 7. 
      During the cultivation period comparative quality control was performed using 
      flow cytometry, gene expression profiling, and functional assays. RESULTS: Both 
      flasks and bags generated mature genetically modified DCs in similar yields. 
      Surface membrane markers, expression profiles, and functional testing results 
      were comparable. The use of a closed-bag system facilitated clinical 
      applicability of genetically modified DCs. CONCLUSIONS: The polyolefin bag-based 
      culture system yields DCs qualitatively and quantitatively comparable to the 
      standard flask preparation. All steps including cryopreservation can be performed 
      in a closed system facilitating standardized, safe, and reproducible preparation 
      of therapeutic cells.
FAU - Macke, Lars
AU  - Macke L
AD  - Department of Molecular Biotechnology, Helmholtz Centre for Infection Research 
      (HZI), Braunschweig, Germany.
FAU - Garritsen, Henk S P
AU  - Garritsen HS
FAU - Meyring, Wilhelm
AU  - Meyring W
FAU - Hannig, Horst
AU  - Hannig H
FAU - Pagelow, Ute
AU  - Pagelow U
FAU - Wormann, Bernhard
AU  - Wormann B
FAU - Piechaczek, Christoph
AU  - Piechaczek C
FAU - Geffers, Robert
AU  - Geffers R
FAU - Rohde, Manfred
AU  - Rohde M
FAU - Lindenmaier, Werner
AU  - Lindenmaier W
FAU - Dittmar, Kurt E J
AU  - Dittmar KE
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20091210
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
RN  - 0 (Antigens, CD)
RN  - 0 (Antigens, Surface)
RN  - 0 (Lipopolysaccharide Receptors)
SB  - IM
MH  - Adult
MH  - Antigens, CD/analysis/genetics
MH  - Antigens, Surface/analysis
MH  - Cell Count
MH  - Cell Culture Techniques/instrumentation/*methods
MH  - Cell Division/physiology
MH  - Cellular Senescence/physiology
MH  - Child
MH  - Dendritic Cells/cytology/*physiology/ultrastructure
MH  - Female
MH  - Gene Transfer Techniques
MH  - Humans
MH  - Leukapheresis/methods
MH  - Leukocytes/cytology/physiology
MH  - Lipopolysaccharide Receptors/analysis/genetics
MH  - Male
MH  - Microscopy, Electron, Scanning
MH  - Middle Aged
MH  - Oligonucleotide Array Sequence Analysis
EDAT- 2009/12/17 06:00
MHDA- 2010/05/06 06:00
CRDT- 2009/12/17 06:00
PHST- 2009/12/17 06:00 [entrez]
PHST- 2009/12/17 06:00 [pubmed]
PHST- 2010/05/06 06:00 [medline]
AID - TRF2520 [pii]
AID - 10.1111/j.1537-2995.2009.02520.x [doi]
PST - ppublish
SO  - Transfusion. 2010 Apr;50(4):843-55. doi: 10.1111/j.1537-2995.2009.02520.x. Epub 
      2009 Dec 10.