Xeno-Free Strategies for Safe Human Mesenchymal Stem/Stromal Cell Expansion: Supplements and Coatings

Abstract

Human mesenchymal stem/stromal cells (hMSCs) have generated great interest in regenerative medicine mainly due to their multidifferentiation potential and immunomodulatory role. Although hMSC can be obtained from different tissues, the number of available cells is always low for clinical applications, thus requiring in vitro expansion. Most of the current protocols for hMSC expansion make use of fetal bovine serum (FBS) as a nutrient-rich supplement. However, regulatory guidelines encourage novel xeno-free alternatives to define safer and standardized protocols for hMSC expansion that preserve their intrinsic therapeutic potential. Since hMSCs are adherent cells, the attachment surface and cell-adhesive components also play a crucial role on their successful expansion. This review focuses on the advantages/disadvantages of FBS-free media and surfaces/coatings that avoid the use of animal serum, overcoming ethical issues and improving the expansion of hMSC for clinical applications in a safe and reproducible way.

Accumulating Transcriptome Drift Precedes Cell Aging in Human Umbilical Cord-Derived Mesenchymal Stromal Cells Serially Cultured to Replicative Senescence

“…The most common source of growth factors to support rapid in vitro expansion of MSCs is animal serum, such as fetal calf serum (FCS). However, FCS is a potential source of pathogens, can elicit immunological reactions in humans, is prone to variable composition, and has a limited production capacity that cannot support commercialized cell therapies . Human blood derivatives, including serum, platelet‐rich plasma, and platelet lysate or fractions thereof, are animal‐free alternatives to FCS but are also subject to source variations that exacerbate the cost of goods .…”

Adipose-derived mesenchymal stem/stromal cells: from the lab bench to the basic concepts for clinical translation

“…Along with their isolation-related qualities, hASC are highly attractive in the medical field, since they do not present surface molecules of class II major histocompatibility complex (MHCII), which allows them to be transplanted across allogenic barriers without eliciting an immune response (Le Blanc et al, 2003;Tse et al, 2003;De Miguel et al, 2012;Huang et al, 2016). If the hASC are to be used in clinics, it is mandatory to replace all traditional culture reagents by defined xeno-free reagents or, alternatively, more recently developed techniques which avoid the use of FBS or other animal sera are to be used (Cimino et al, 2017).…”

The Culture Medium as a Key Factor of the Mammalian Cell Cultivation

“…Некоторые авторы также выделяют среды, не содержащие чужеродных компонентов (xeno-free media или animal-derived component-free media), в состав которых входят только соединения человеческого, но не животного происхождения (например, сывороточный альбумин человека) [10]. При этом среды, не содержащие компоненты животного происхождения, могут включать бактериальные, дрожжевые гидролизаты или растительные экстракты [1,5].…”

“…В случаях, когда применение сыворотки является нежелательным, исследователи могут использовать так называемые заменители сыворотки [8,27]. Они включают экстракты сыворотки, экстракты тканей или гидролизаты [10], в том числе лизаты тромбоцитов факторы роста [28,29], гормоны, белки-переносчики (альбумин и трансферрин), липиды, металлы, витамины, полиамины и восстановители (2-меркапоэтанол, α-тиоглицерин, восстановленный глутатион) [30,31]. Количество комбинаций этих соединений практически бесконечно, при этом они, как правило, взаимодействуют друг с другом.…”