Human Platelet Lysate Improves Bone Forming Potential of Human Progenitor Cells Expanded in Microcarrier-Based Dynamic Culture

Abstract

Xenogeneic‐free media are required for translating advanced therapeutic medicinal products to the clinics. In addition, process efficiency is crucial for ensuring cost efficiency, especially when considering large‐scale production of mesenchymal stem cells (MSCs). Human platelet lysate (HPL) has been increasingly adopted as an alternative for fetal bovine serum (FBS) for MSCs. However, its therapeutic and regenerative potential in vivo is largely unexplored. Herein, we compare the effects of FBS and HPL supplementation for a scalable, microcarrier‐based dynamic expansion of human periosteum‐derived cells (hPDCs) while assessing their bone forming capacity by subcutaneous implantation in small animal model. We observed that HPL resulted in faster cell proliferation with a total fold increase of 5.2 ± 0.61 in comparison to 2.7 ± 02.22‐fold in FBS. Cell viability and trilineage differentiation capability were maintained by HPL, although a suppression of adipogenic differentiation potential was observed. Differences in mRNA expression profiles were also observed between the two on several markers. When implanted, we observed a significant difference between the bone forming capacity of cells expanded in FBS and HPL, with HPL supplementation resulting in almost three times more mineralized tissue within calcium phosphate scaffolds. FBS‐expanded cells resulted in a fibrous tissue structure, whereas HPL resulted in mineralized tissue formation, which can be classified as newly formed bone, verified by μCT and histological analysis. We also observed the presence of blood vessels in our explants. In conclusion, we suggest that replacing FBS with HPL in bioreactor‐based expansion of hPDCs is an optimal solution that increases expansion efficiency along with promoting bone forming capacity of these cells. Stem Cells Translational Medicine 2019;8:810&821

Chitosan-Hydrogel Polymeric Scaffold Acts as an Independent Primary Inducer of Osteogenic Differentiation in Human Mesenchymal Stromal Cells

“…Similar results were reported in different studies, and this is a turning point for the application of a scaffold with a peculiar chemical composition [46][47][48]. In fact, the structure and the materials must not interact with the vital function of the cells in order to be suitable for regenerative medicine application [49].…”

Large-Scale Expansion of Human Mesenchymal Stem Cells

“…Several types of spinner flasks, including the Bellco spinner flask [ 18 , 24 , 25 , 30 , 31 , 33 ], Techne spinner flask [ 7 , 26 ], Corning spinner flask [ 29 ], and Cellspin spinning bottle [ 16 ] with capacity ranging from 100 ml to 125 ml, have been used for large-scale expansion of MSCs. Microcarriers were used to provide the growth surface for cell proliferation.…”

“…All three studies reported that FBS was inferior compared to HPL and defined medium in promoting MSC proliferation. Instead of FBS, seven studies used 5%, 8%, or 10% (v/v) HPL [17,[19][20][21][22][23][24], one study used 5% (v/v) Ultragrow™ [25], and another one used 15% (v/v) AB human serum as supplement [26]. Defined culture mediums, i.e., MesenCult™-XF medium [27][28][29], StemPro® MSC SFM XenoFree medium [30][31][32][33], Corning® stemgro® hMSC medium [29], and PRIME-XV™ SFM medium [18], were used in seven studies ( Table 2).…”

Three dimensional microcarrier system in mesenchymal stem cell culture: a systematic review

“…Under xenogenic-free culture conditions, Gupta et al reported that HPL resulted in faster cell proliferation by 5.2 ± 0.61-fold in comparison to 2.7 ± 02.22-fold in FBS [60]. In addition, an automated serum-free, microcarrier culture system was established.…”