Human platelet lysate allows expansion and clinical grade production of mesenchymal stromal cells from small samples of bone marrow aspirates or marrow filter washouts

Abstract

We compared two protocols for the expansion of human mesenchymal stromal cells (hMSCs) starting from diagnostic samples of BM aspirates (2-5 ml) or using the remnants in the bag and filter at the end of the BM infusions. The protocols differed in the presence of either 10% fetal bovine serum (FBS) or 5% platelet lysate (PL). We obtained a significantly (P ¼ 0.02) better expansion with PL, obtaining a median 1010-fold compared to 198-fold with a selected batch of FBS and in fewer days (29.8 in PL versus 41.4 in FBS). Overall, we recovered a variable number from 54.8 Â 10 6 to 365 Â 10 6 hMSCs in PL versus a variable number from 2.7 Â 10 6 to 31 Â 10 6 in FBS. No difference could be found in terms of gross morphology, differentiation potential, surface markers and immunological properties (inhibition of allogeneic PHA response and mixed lymphocyte reaction) of cells expanded with PL or FBS. The preparations were found within the range of acceptability for all the quality control criteria. Due to the clinical grade nature of the PL and the reproducibility of separate preparations, we propose this method to obtain hMSCs even from minute amounts of BM cells.

Inactivated human platelet lysate with psoralen: a new perspective for mesenchymal stromal cell production in Good Manufacturing Practice conditions

“…Moreover, it represents a good GMP-compliant alternative to animal serum for MSC clinical production confirming the recent data reported in the literature (18,36,37).…”

Autologous Mesenchymal Stromal Cells and Kidney Transplantation

“…MSCs were isolated and ex vivo expanded according to Good-ManufacturingPractice procedures (Cell-Therapy Laboratory "G. Lanzani", Ospedali Riuniti di Bergamo, authorization no. aM-189/2008 Agenzia Italiana del Farmaco, AIFA) (23,24). On day 7 after kidney transplant, autologous MSCs were administered intravenously (1.7 ϫ 10 6 cells and 2.0 ϫ 10 6 cells per kg body weight, respectively) after premedication with chlorphenamine and acetaminophen.…”

“…MSCs were processed and cultured as previously reported (23,24). In brief, bone marrow aspirates were collected and nucleated cells were plated at 500,000 cells per cm 2 in minimum essential medium-␣ in the presence of 5% human platelet lysate.…”

Adipocyte Derived Mesenchymal Stromal Cell and Platelet Lysate: Ideal Cell and Supplement for the Treatment of Immune-Inflammatory Diseases?

“…In brief, filters were washed with 200 mL PBS (Gibco, Invitrogen Corp., USA); cells were centrifuged at 400 g 10 min, counted and cultured as stated above [14].…”