A new method to confirm the absence of human and animal serum in mesenchymal stem cell culture media

Abstract

Mesenchymal stem cells are an ideal source for regenerative medicine. For clinical use, cell culture should be done at stable conditions, thus the use of serum should be avoided because of the batch-to-batch variations of serum. Although several kinds of serum-free media are available, a method to confirm whether they contain serum has not been established yet. During studies on effect of adipocyte mesenchymal stem cells (Ad-MSCs) on pain using a human pain gene array, we noticed that BDKRB1 gene was constantly upregulated when serum was used in the culture medium. In this study, we attempted to establish further the potential of this gene as a new marker indicative of the presence of serum in media. Using a real-time quantitative PCR gene array screening containing 84 functional genes, we verified BDKRB1 as a specific gene upregulated in the presence of serum. The expression of BDKRB1 in Ad-MSCs was induced not only by bovine serum but also by human serum. The BDKRB1 expression was induced even when Ad-MSCs was cultured with 0.1% serum in the medium. We concluded that BDKRB1 is a valuable marker to detect traces of both human and animal serum in Ad-MSCs cultures. Our study provides a new method to confirm the absence of serum in media and ensure a stable cell culture condition.

Three dimensional microcarrier system in mesenchymal stem cell culture: a systematic review

“…The variation from each batches of serum further affect the quality of the up-scaled product by different sources of origin, brands, and present of unidentified risk of contamination. In this case, one of the solutions to minimise this variations is the adaptation of serum free media (SFM) in MSC culture as mentioned by Ota et al [68]. Aside from cell yield variations, the downstream harvesting approaches still require optimisation to improve cell recovery; in fact, MSC differentiation efficiency in 3D system remains uncertain and the mechanism is still not well-studied.…”

Comparative analysis of mesenchymal stem cells cultivated in serum free media

“…4 ). Neu5Gc was used as the marker of the animal serum-derived substance 14 , and BDKRB1 as the marker of the human serum-derived substance 48 . All tested SFM did not contain animal serum-derived substances, but BDKRB1, human serum-derived substance marker, showed significantly high expression in SFM B (Supplementary Fig.…”

Development of Low-Serum Culture Media for the in Vitro Cultivation of Theileria annulata S15 Cell Line

“…Serum may contain contaminants such as mycoplasma, viruses, and prion agents that cause BSE in humans and animals ( 2 ). Furthermore, the chemical composition of serum is variable across different batches ( 3 ).…”