Human platelet lysate supports the formation of robust human periodontal ligament cell sheets

Abstract

The use of stem cell-derived sheets has become increasingly common in a wide variety of biomedical applications. Although substantial evidence has demonstrated that human platelet lysate (PL) can be used for therapeutic cell expansion, either as a substitute for or as a supplement to xenogeneic fetal bovine serum (FBS), its impact on cell sheet production remains largely unexplored. In this study, we manufactured periodontal ligament stem cell (PDLSC) sheets in vitro by incubating PDLSCs in sheet-induction media supplemented with various ratios of PL and FBS, i.e. 10% PL without FBS, 7.5% PL + 2.5% FBS, 5% PL + 5% FBS, 2.5% PL + 7.5% FBS or 10% FBS without PL. Cultures with the addition of all the designed supplements led to successful cell sheet production. In addition, all the resultant cellular materials exhibited similar expression profiles of matrix-related genes and proteins, such as collagen I, fibronectin and integrin β1. Interestingly, the cell components within sheets generated by media containing both PL and FBS exhibited improved osteogenic potential. Following in vivo transplantation, all sheets supported significant new bone formation. Our data suggest that robust PDLSC sheets can be produced by applying PL as either an alternative or an adjuvant to FBS. Further examination of the relevant influences of human PL that benefit cell behaviour and matrix production will pave the way towards optimized and standardized conditions for cell sheet production.

The effects of conditioned media generated by polarized macrophages on the cellular behaviours of bone marrow mesenchymal stem cells

“…Based on qRT‐PCR, immunofluorescent staining of transverse images and Western blotting, we found that CM1‐ and CM2‐based inductive media significantly elevated the abundance of fibronectin, COL‐1 and integrin β1 protein (Figs C–E and A–E). In our previous study, periodontal ligament stem cell (PDLSC) sheets were manufactured in vitro by incubating cells in sheet‐induction media supplemented with various ratios of human platelet lysate (PL) and xenogeneic FBS, and immunohistochemical staining suggested that all resultant cellular materials displayed similar protein profiles of fibronectin, COL‐1 and integrin β1 . Thus, it appears that immunohistochemical staining of lateral sections is not necessary to evaluate the protein profiles of stem cell sheets.…”

“…Haematoxylin and eosin staining was performed, and the stained sheets were analysed using previously reported methods . Briefly, cell sheets were fixed in 4% paraformaldehyde for 12 hrs.…”

Human periodontal ligament stem cells on calcium phosphate scaffold delivering platelet lysate to enhance bone regeneration

“…25,26 However, a literature search revealed only one report that investigated the effect of HPL on hPDLSCs. 27 In that study, HPL was added into the culture medium, without being delivered via a scaffold. The HPL beneted the matrix production by the hPDLSCs, the osteogenic gene expressions were upregulated, and the mineralization of hPDLSCs was increased.…”

Influence of Human Platelet Lysate on Extracellular Matrix Deposition and Cellular Characteristics in Adipose-Derived Stem Cell Sheets

“…HPL was suggested to alter the ASC behavior and function via multiple signaling pathways (Kakudo et al, 2019). A previous study showed that HPL can benefit cell behavior and ECM production, thus optimizing the conditions for cell sheet production in periodontal ligament stem cells (Tian et al, 2018). Since the application of HPL in fabricating ASC sheets had not been reported in the literature, it was therefore intriguing to explore the effect of replacing FBS with HPL on the microenvironment fabricated within ASC sheets.…”