Isolation, expansion and characterization of bone marrow-derived mesenchymal stromal cells in serum-free conditions

Abstract

Bone marrow-derived mesenchymal stromal cells (BM-MSCs) heralded a new beginning for regenerative medicine and generated tremendous interest as the most promising source for therapeutic application. Most cell therapies require stringent regulatory compliance and prefer the use of serum-free media (SFM) or xeno-free media (XFM) for the MSC production process, starting from the isolation onwards. Here, we report on serum-free isolation and expansion of MSCs and compare them with cells grown in conventional fetal bovine serum (FBS)-containing media as a control. The isolation, proliferation and morphology analysis demonstrated significant differences between MSCs cultured in various SFM/XFM in addition to their difference with FBS controls. BD Mosaic™ Mesenchymal Stem Cell Serum-Free media (BD-SFM) and Mesencult-XF (MSX) supported the isolation, sequential passaging, tri-lineage differentiation potential and acceptable surface marker expression profile of BM-MSCs. Further, MSCs cultured in SFM showed higher immune suppression and hypo-immunogenicity properties, making them an ideal candidate for allogeneic cell therapy. Although cells cultured in control media have a significantly higher proliferation rate, BM-MSCs cultured in BD-SFM or MSX media are the preferred choice to meet regulatory requirements as they do not contain bovine serum. While BM-MSCs cultured in BD-SFM and MSX media adhered to all MSC characteristics, in the case of few parameters, the performance of cells cultured in BD-SFM was superior to that of MSX media. Pre-clinical safety and efficiency studies are required before qualifying SFM or XFM media-derived MSCs for therapeutic applications.

Evaluation of the Secretome Profile and Functional Characteristics of Human Bone Marrow Mesenchymal Stromal Cells-Derived Conditioned Medium Suggest Potential for Skin Rejuvenation

“…The bioactive factors secreted by MSC may eventually fill this niche because of their potential to repair skin damage and aid in skin rejuvenation. Amongst the latest developments in the cosmetic industry, use of biotechnology-derived ingredients, nutritional regimens, fibroblast or stem cell-based products are becoming increasingly popular, particularly for the aging population [13]. The unique characteristics of MSCs unraveled over the past decades, are not only being exploited for the rapidly growing field of rege-nerative medicine, but also provide a promising platform for generation of novel cosmetics.…”

“…We had earlier reported and published the development and characterization of a pooled, bone marrow derived MSC population, obtained from healthy, consenting donors, that were expanded in serum-containing or serum-free medium in a reproducible manner in a GMP compliant facility [11] [12] [13].…”

Venous Blood Derivatives as FBS-Substitutes for Mesenchymal Stem Cells: A Systematic Scoping Review

“…The recent literature has been investigated FBS substitutes for MSC ex vivo expansion aiming to eliminated the risks inherent to xenogeneic agents for clinical translation of regenerative therapies (103,104). In this systematic scoping review, the studies evaluating VBD as FBS substitutes were summarized.…”

Fetal bovine serum (FBS): Past – present – future

“…With an almost exponentially increasing number of newly identified or generated cell lines/ types every year (Freshney, 2005), more cell-specific media are needed. Non-specific media often lead to slower growth compared to FBS containing media (Chowdhury et al, 2012;Gottipamula et al, 2014). Furthermore, the substitution of serum with defined components requires not only extensive know-how and experience, but also a wide range of biochemical substances of high purity grade, the commercial availability of which is not always guaranteed or optimal.…”