Evaluation of platelet lysate as a substitute for FBS in explant and enzymatic isolation methods of human umbilical cord MSCs

Abstract

Mesenchymal stem cells (MSCs) have immense potential for cell-based therapy of acute and chronic pathological conditions. MSC transplantation for cell-based therapy requires a substantial number of cells in the range of 0.5–2.5 × 106 cells/kg body weight of an individual. A prolific source of MSCs followed by in vitro propagation is therefore an absolute prerequisite for clinical applications. Umbilical cord tissue (UCT) is an abundantly available prolific source of MSC that are fetal in nature and have higher potential for ex-vivo expansion. However, the ex-vivo expansion of MSCs using a xenogeneic supplement such as fetal bovine serum (FBS) carries the risk of transmission of zoonotic infections and immunological reactions. We used platelet lysate (PL) as a xeno-free, allogeneic replacement for FBS and compared the biological and functional characteristics of MSC processed and expanded with PL and FBS by explant and enzymatic method. UCT-MSCs expanded using PL displayed typical immunophenotype, plasticity, immunomodulatory property and chromosomal stability. PL supplementation also showed 2-fold increase in MSC yield from explant culture with improved immunomodulatory activity as compared to enzymatically dissociated cultures. In conclusion, PL from expired platelets is a viable alternative to FBS for generating clinically relevant numbers of MSC from explant cultures over enzymatic method.

Autologous adipose-derived stem cells for the treatment of complex cryptoglandular perianal fistula: A randomized clinical trial with long-term follow-up

“…Unlike in other studies, here ASCs were expanded using human platelet lysate in the culture media, which may have the advantage of avoiding the use of animal‐derived factors. Human platelet lysate is widely used to grow MSCs, and many studies have demonstrated that this confers a degree of safety that is at least equivalent to that offered by fetal bovine serum . Indeed, some studies have suggested that the use of platelet lysate may increase cell yield and shorten the time for ex vivo expansion without epigenetic alterations …”

“…Human platelet lysate is widely used to grow MSCs, [23][24][25] and many studies have demonstrated that this confers a degree of safety that is at least equivalent to that offered by fetal bovine serum. 26,27 Indeed, some studies have suggested that the use of platelet lysate may increase cell yield and shorten the time for ex vivo expansion without epigenetic alterations.…”

“…26,27 Indeed, some studies have suggested that the use of platelet lysate may increase cell yield and shorten the time for ex vivo expansion without epigenetic alterations. 25,27 One of the issues regarding the use of somatic cell medicinal products has been the selection of the cell dose. In general, decisions regarding dose levels were based on studies performed in animals, the absence of AEs, and the number of cells that can be generated in a particular period.…”

Maternal Serum, an Isolation and Expansion Tool for Umbilical Cord Matrix Mesenchymal Stromal Cells

“…Recently, it has been highlighted that there is a need to find alternatives to FBS for use as a media supplement for MSC culture, due to the risk of transmitting pathogens that may cause viral infections and bacterial infections, transmission of prions, inflammatory reactions due to contamination with bovine proteins, and ethical issues with collection of animal products [11,12,15,17]. Alternatives have been considered: MSCs have been cultured successfully in autologous [18,19] and non-autologous [20,21] human serum, human platelet lysate [12,17,20], autologous cord blood serum [20,22], and autologous cord blood platelet rich plasma [23,24]. Serum is collected from whole blood after it is left for a specific set time to clot and is then subsequently centrifuged [25,26].…”

“…Platelet rich plasma is produced when the platelet pellet is re-suspended in the plasma after centrifugation [27,24]. Platelet lysate is produced when the platelet rich plasma is taken through a number of freeze thaw cycles [25,28,29].…”