Feasibility and Efficiency of Human Bone Marrow Stromal Cell Culture with Allogeneic Platelet Lysate‐Supplementation for Cell Therapy against Stroke

Abstract

Currently, there is increasing interest in human bone marrow stromal cells (hBMSCs) as regeneration therapy against cerebral stroke. The aim of the present study was to evaluate the feasibility and validity of hBMSC cultures with allogeneic platelet lysates (PLs). Platelet concentrates (PC) were harvested from healthy volunteers and made into single donor-derived PL (sPL). The PL mixtures (mPL) were made from three different sPL. Some growth factors and platelet cell surface antigens were detected by enzyme-linked immunosorbent assay (ELISA). The hBMSCs cultured with 10% PL were analyzed for their proliferative potential, surface markers, and karyotypes. The cells were incubated with superparamagnetic iron oxide (SPIO) agents and injected into a pig brain. MRI and histological analysis were performed. Consequently, nine lots of sPL and three mPL were prepared. ELISA analysis showed that PL contained adequate growth factors and a particle of platelet surface antigens. Cell proliferation capacity of PLs was equivalent to or higher than that of fetal calf serum (FCS). No contradiction in cell surface markers and no chromosomal aberrations were found. The MRI detected the distribution of SPIO-labeled hBMSCs in the pig brain. In summary, the hBMSCs cultured with allogeneic PL are suitable for cell therapy against stroke.

A specific protocol of autologous bone marrow concentrate and platelet products versus exercise therapy for symptomatic knee osteoarthritis: a randomized controlled trial with 2 year follow-up

“…The addition of autologous PRP and PL for the post-injection is primarily to aid in the proliferation of both native MSCs and those contained in the BMC. The benefits of PL as a culture medium for MSCs and its effects on cellular proliferation have also been widely reported [32].…”

Novel therapeutic approaches in utilizing platelet lysate in regenerative medicine: Are we ready for clinical use?

“…[Color figure can be viewed at wileyonlinelibrary.com] T A B L E 2 Details of the preparation techniques of platelet lysate; the type of use, platelet source, platelet count and preparation method in clinical applications, If reported, is listed. At first, animal studies are presented and then the human studies : phosphate-buffered saline Huang et al, 2017;C. Tan & Shichinohe, 2016;von Bonin et al, 2009…”

Research on advanced intervention using novel bone marrOW stem cell (RAINBOW): a study protocol for a phase I, open-label, uncontrolled, dose-response trial of autologous bone marrow stromal cell transplantation in patients with acute ischemic stroke

“…BMMNCs are isolated via density-gradient centrifugation with Ficoll-Hypaque® (Pharmacia, Uppsala, Sweden), and 1 × 10 7 cells are plated in a 175 cm 2 uncoated flask (Easy Flask 159,910; Nunc) with 25 mL of αMEM with 10% hPL derived from healthy volunteers and 40 μg/mL of gentamicin sulfate. About the preparation of hPL, our previous report about safety and efficiency of hPL should be referred to [ 4 ]. After 24 h, nonadherent cells are removed by changing the medium.…”

“…BMSCs originate from bone marrow mononuclear cells (BMMNCs). The floating cells are removed from BMMNCs, and only cells adhering to the floor of the flask are cultured for several weeks to obtain BMSCs [ 4 ]. A subpopulation of BMSCs, also termed mesenchymal stem cells, can differentiate into bone, cartilage, and fat.…”