Human platelet lysate (hPL) is rich in growth factors (GF) and nutritive elements
and represents a powerful xeno-free alternative to fetal bovine serum (FBS) notably
for mesenchymal stem cell (hMSC) proliferation. However, there is a large variability
in hPL sources and production protocols, resulting in discrepancies in product quality,
low management of product safety and poor batch-to-batch standardization. We describe
here the development and the characterization of a standardized hPL prepared from
outdated transfusional grade screened normal human donor platelet concentrates (PCs),
manufactured on an industrial scale (batch sizes of 10 L; 250 donors) and following
a highly qualified process (clean room, trained operators, validated aseptic filtration).
PCs were frozen at −80°C and thawed at +4°C to lyse platelets. Cell debris were removed
by centrifugation and the supernatant (hPL) was recovered. Clinical grade 10L batches
of aseptic filtered hPL were characterized. First, we showed that hPL prepared from
a limited number of donors displayed a variability in terms of GF contents. On the
contrary, we observed a robust standardization between 10L-industrial batches of hPL
in terms of GF contents (bFGF, EGF, VEGF, PDGF-AB, TGF-beta1 and IGF-1), biochemical
analyses (total proteins, albumin, fibrinogen and iron) and efficacy on bone marrow
(BM)-hMSC proliferation. Secondly, we compared expansion and functional characteristics
of BM-hMSCs grown in clinical grade hPL versus MSC-screened FBS batch. We showed a reproducible increase in cell growth kinetics
using hPL, a maintenance of BM-hMSC clonogenic potential and membrane marker expression
(with however a strong overexpression of CD90). We observed a similar adipogenic and
osteogenic differentiation potential and finally that immunosuppressive properties
of BM-hMSCs (inhibition of T-cell proliferation) cultivated in parallel in both conditions
remained also identical. Finally, we documented the stability over time of hPL stored
at −80°C and −20°C. In conclusion, we demonstrated the feasibility to use a standardized,
efficient and clinical grade hPL for research and cell therapy applications.
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to CytotherapyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect