Expansion of adipose tissue mesenchymal stromal progenitors in serum-free medium supplemented with virally inactivated allogeneic human platelet lysate.
BACKGROUND
Single-donor or pooled platelet lysates (PL) can substitute for fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) expansion. However, for clinical applications of MSCs, the use of virally inactivated PL would be desirable. Recently, we have developed a solvent/detergent (S/D)-treated human PL preparation (S/D-PL) rich in growth factors. The capacity to use this virally inactivated preparation for MSC expansion needs to be evaluated.


METHODS
Platelet concentrates were treated by S/D (1% tri-n-butyl phosphate and 1% Triton X-45), extracted by oil, purified by C18 hydrophobic interaction chromatography, and sterile filtered. S/D-PL was compared to FBS as a medium supplement (10% vol/vol) for isolating, maintaining, and expanding adipose tissue-derived MSCs (AT-MSCs). Cell morphology; proliferation kinetics; immunophenotype; differentiation capacity toward the chondrogenic, osteogenic, and osteogenic lineages; and cytokine antibody array were assessed.


RESULTS
AT-MSCs had a typical spindle morphology and proliferated in S/D-PL at least as well as in FBS. Immunophenotype at Passage 7 was characteristic of MSCs and similar for both culture conditions. Differentiation capacity into the three lineages was maintained and chondrogenesis was enhanced by S/D-PL. In a 120 human cytokine antibody array analysis, 73 cytokines were detected in S/D-PL, including 22 with a concentration higher than in FBS.


CONCLUSIONS
S/D-PL is an alternative to FBS for AT-MSC maintenance and expansion, does not compromise the differentiation capacity nor the immunophenotype, and may accelerate chondrogenesis. S/D-PL protocols for MSC clinical scale-up may represent a major step toward challenging new use in stem cell therapies.
Graduate Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan.
Daniel Tzu-Bi
DT
Shih
Jung-Cheu
JC
Chen
Wan-Yu
WY
Chen
Ya-Po
YP
Kuo
Chen-Yao
CY
Su
Thierry
T
Burnouf
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D001792:Blood Platelets / Q000737:chemistry*; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects
D016895:Culture Media, Serum-Free