Lyophilized human platelet lysate: manufacturing, quality control, and application

Area of Science:

• Biotechnology
• Cell Biology
• Regenerative Medicine

Background:

• Platelet concentrates (PCs) are often discarded due to short shelf life, leading to waste.
• Human platelet lysate (HPL) from expired PCs is a promising, ethical alternative to animal serum in cell culture.
• Current HPL use is limited by batch variability and poor storage stability.

Purpose of the Study:

• To develop a lyophilization protocol for HPL to improve its stability and applicability.
• To evaluate the impact of lyophilization on HPL's quality control parameters and growth factor content.
• To assess the functionality of lyophilized HPL (L-HPL) in supporting human mesenchymal stromal cell (hMSC) culture and differentiation.

Main Methods:

• Paired comparison of six HPL and L-HPL batches for pH, protein, osmolality, and electrolyte concentrations.
• Quantification of 11 growth factors and cytokines in HPL versus L-HPL.
• Assessment of hMSC yield, proliferation, viability, and trilineage differentiation potential in media supplemented with HPL or L-HPL.

Main Results:

• L-HPL showed comparable pH, osmolality, and potassium levels to HPL, with slightly lower total protein, sodium, and chloride.
• Growth factor and cytokine concentrations were consistent between HPL and L-HPL.
• hMSCs cultured with L-HPL exhibited similar cell yield, division cycles, and viability compared to those cultured with HPL.
• Osteogenic differentiation capacity was slightly enhanced in cells cultured with L-HPL, while adipogenic and chondrogenic potentials remained unchanged.

Conclusions:

• A robust protocol for producing stable, applicable L-HPL was successfully established.
• L-HPL demonstrates comparable quality control parameters, growth factor profiles, and hMSC functionality to standard HPL.
• L-HPL serves as a suitable, stable, and ethical cell culture supplement, offering a viable alternative to animal-derived sera in research and therapeutic applications.