Human Pluripotent Stem Cells (PSCs), including Embryonic Stem Cells (ESCs) and induced
Pluripotent Stem Cells (iPSCs), have the remarkable potential to self-renew and develop into
various cell lineages. Human Mesenchymal Stem Cells (MSCs) or multipotent stem cells that
are present in various organs can self-renew and differentiate into multiple mesenchymal lineages.
Both human PSCs and MSCs hold great promise in cell-based therapies, disease modeling, drug
discovery, and regenerative medicine. Human stem cells must be cultured under the optimal conditions
to use them in transplantology. Therefore, researchers must ensure the sterility of human stem
cell lines. Bacterial contamination is a common problem in laboratories and major precautions are
required to detect the types of microorganisms, and to eliminate and prevent contamination in cell
cultures. Stem cell culture media usually contain antibiotics and antimycotics such as penicillin-
streptomycin (pen-strep), gentamicin, and amphotericin B (AmB) to avoid bacterial, fungal,
and yeast contaminants. Numerous publications recognized the serious effect of antibiotics and antimycotics
on in vitro properties of human stem cells, including proliferation, differentiation, survival,
and genetic instability. This review study aimed to understand the impact of routinely used
antibiotics and antimycotics such as pen-strep, gentamicin, and AmB on viability, proliferation,
and functional properties (differentiation and pluripotency) of human PSCs and MSCs.