Development of standard protocol for the cGMP production of human mesenchymal stem/stromal cells from different source tissues

Abstract

Background & Aim Mesenchymal stem/stromal cells (MSC) hold great potential for tissue engineering and regenerative medicine because of their ability for self-renewal and differentiation into tissue-specific cells such as osteoblasts, chondrocytes, and adipocytes. MSCs are important pieces in the process of tissue development, maintenance and repair, producing secretory factors that support engraftment and trophic functions. For that reason, they are currently being used in hundreds of clinical trials as a promising treatment option for a variety of clinical conditions, with an increasing evidence of therapeutic efficacy. MSC constitute a heterogeneous subset of stromal regenerative cells which can be harvested from several adult tissues. Initially MSC were identified in bone marrow but currently they can be harvested from several autologous and allogeneic sources, including adipose tissue, peripheral blood, lungs, marrow spaces of long bone, synovial fluids, periodontal ligament, dental pulp and muscle. In addition, MSCs are also obtained from placenta, umbilical cord and cord blood. Notably, MSCs obtained from various sources differ in their biological characteristics, CFU-F efficiency, specific markers, multi-lineage differentiation as well as paracrine functions which may determine their different clinical applications. The use of MSC from different sources together with the fact that most laboratories use their own standardized protocols for cell expansion results in a clear variability in cell preparations between labs which can lead to variability in the outcome of experiments and clinical trials across different laboratories and institutions. Investigations have attempted to address this issue of unpredictable outcomes by seeking to establish standard practices for the isolation, characterization, and maintenance of cells in culture. The development of uniform protocols for the expansion and preparation of MSCs is crucial to improve comparison of results, for the exploitation of MSC clinical potential and for a successful translation of approaches to the clinic. Methods, Results & Conclusion Here we will present the development of a standardized protocol for a consistent and efficient cGMP production of clinically relevant numbers of MSCs obtained from different source tissues, using MSC NutriStem® XF Basal Medium supplemented with PLTGold®, a fully xenogeneic free human platelet lysate.