Ex vivo expansion of umbilical cord blood CD34 cells in a closed system: a multicentric study.
OBJECTIVE
The Dideco 'Pluricell System' is a CE-marked medical device allowing haematopoietic stem cell (HSC) expansion. It comprises a kit of cGMP cytokines and reagents, a closed-cell expansion chamber and a cell-washing set. We tested the system in a multicentric study by expanding CD34(+) cells from eight fresh umbilical cord blood (UCB) samples.


METHODS
During culture, the mean nucleated cell (NC) count, the mean CD34(+) cell count, fold expansion, viability and apoptosis were measured. Clonogenic assays and immunophenotypical characterization were performed on days 0, 7 and 12. On the expanded cellular product, in three cases cell genotyping, endotoxin level and mycoplasma detection (by polymerase chain reaction) were performed.


RESULTS
The mean CD34(+) cell expansion on days 7 and 12 was sevenfold and 12-fold respectively and the mean NC expansion was 69-fold and 180-fold. The mean NC viability on day 12 was 96.9% (94.4-99.1). After 12 days, granulocyte-macrophage colony-forming units (GM-CFU) showed a 20-fold increase: a slight increase in CD34(+) cell apoptosis was observed during culture. In all of three cases neither chromosomal alterations nor mycoplasma contamination was detected. No significant endotoxin levels were detected after expansion.


CONCLUSIONS
The device allows the ex vivo expansion of NC and CD34(+) cells in a closed system. The expanded cellular product is a mixture of progenitors (CD34(+) cells) and differentiated (mainly myeloid and megakaryocytic) cells. To reduce cell apoptosis, more frequent cell feeding during culture should be tested.
Astori
G
G
DIDECO srl Mirandola, MO, Italy.
Larghero
J
J
Bonfini
T
T
Giancola
R
R
Di Riti
M
M
Rodriguez
L
L
Rodriguez
M
M
Mambrini
G
G
Bigi
L
L
Lacone
A
A
Marolleau
J P
JP
Panzani
I
I
Garcia
J
J
Querol
S
S
D018952:Antigens, CD34; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D005312:Fetal Blood / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D011933:Reagent Kits, Diagnostic
D018952:Antigens, CD34; D011933:Reagent Kits, Diagnostic