Development and Characterization of a Clinically Compliant Xeno-Free Culture Medium in Good Manufacturing Practice for Human Multipotent Mesenchymal Stem Cells

Abstract

Human multipotent mesenchymal stem cell (MSC) therapies are currently being tested in clinical trials for Crohn's disease, multiple sclerosis, graft-versus-host disease, type 1 diabetes, bone fractures, cartilage damage, and cardiac diseases. Despite remarkable progress in clinical trials, most applications still use traditional culture media containing fetal bovine serum or serum-free media that contain serum albumin, insulin, and transferrin. The ill-defined and variable nature of traditional culture media remains a challenge and has created a need for better defined xeno-free culture media to meet the regulatory and long-term safety requirements for cell-based therapies. We developed and tested a serum-free and xeno-free culture medium (SFM-XF) using human bone marrow-and adipose-derived MSCs by investigating primary cell isolation, multiple passage expansion, mesoderm differentiation, cellular phenotype, and gene expression analysis, which are critical for complying with translation to cell therapy. Human MSCs expanded in SFM-XF showed continual propagation, with an expected phenotype and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages similar to that of MSCs expanded in traditional serum-containing culture medium (SCM). To monitor global gene expression, the transcriptomes of bone marrow-derived MSCs expanded in SFM-XF and SCM were compared, revealing relatively similar expression profiles. In addition, the SFM-XF supported the isolation and propagation of human MSCs from primary human marrow aspirates, ensuring that these methods and reagents are compatible for translation to therapy. The SFM-XF culture system allows better expansion and multipotentiality of MSCs and serves as a preferred alternative to serum-containing media for the production of large scale, functionally competent MSCs for future clinical applications. STEM CELLS TRANSLATIONAL MEDI-CINE 2012;1:750 -758

Different Culture Conditions Modulate the Immunological Properties of Adipose Stem Cells

“…In fact, several reports have been published to fully characterize ASCs and to develop safe and efficient in vitro culture protocols for clinical cell therapies [30][31][32][33][34]. Completely defined xeno-free and serum-free (XF/SF) isolation and expansion protocols were recently developed for ASCs by our team [30] to replace traditional fetal bovine serum (FBS)-based culturing protocols, which should be avoided in clinical treatments because of safety concerns [35,36].…”

Induced Pluripotent and Mesenchymal Stem Cells as a Promising Tool for Articular Cartilage Regeneration

“…Especially, the nonhuman sialic acid Neu5Gc molecules secreted by feeder cells may contribute to the rejection [81]. Human bone marrow-and adiposederived MSCs were effectively expanded in serum-free and xeno-free culture medium (SFM-XF), and their capacity to differentiate into adipogenic, chondrogenic or osteogenic lineages was preserved [82]. Animal-free hESC pluripotency can be maintained by their encapsulation in calcium alginate hydrogels and their growth in basic medium is retained [83].…”

Characterization of Human AB Serum for Mesenchymal Stromal Cell Expansion

“…Another approach described in the literature involves the use of chemically defined serum-free media for MSC culture [26][27][28][29], therefore reducing batch-to-batch variability. Although a promising alternative, this culture does not maintain an expansion pattern similar to that obtained in cultures supplemented with serum [26,27].…”