Human mesenchymal stem cell expansion on laminin-521 in serum-free and xeno-free culture conditions.

Abstract

Laminin-521 (LN521) is a crucial adhesion protein found in natural stem cell niches and plays an important role in maintaining human pluripotent stem cell (PSC) properties. This study aimed to investigate the effects of LN521 on human umbilical cord-derived mesenchymal stem cell (UC-MSC) characteristics in Serum-free and Xeno-free culture conditions as a step toward clinical application. In our experiment isolated UC-MSC via explant method were expanded as a homogeneous monolayer and morphologically, presented typical MSC-like morphology (spindle-shaped) from passage three to six when cultured on either LN521 or CELLstart™. Almost, 90% confluency was reached after 4 days of culture with an EI of approximately 11.2 with no statistically significant differences on LN521 and CELLstart™ in all six passages. Phenotypic characterization of UC-MSC cultured on LN521 or CELLstart™ using flow cytometry, along with the expression of the same biomarkers in gene level analyzed by quantitative reversed transcription revealed identical CD73, CD90, CD105, CD34, CD45, CD19, CD14, and HLA-DR expression pattern at passages three and six in both LN521 and CELLstart™. Moreover, UC-MSC cultured in the presence of LN521 and CELLstart™ showed the same adipogenesis, chondrogenesis and osteogenesis differentiation potential, and normal chromosome structure highlighting genetic stability. Ultimately, LN521 is comparable to CELLstart™ in supporting UC-MSC expansion and maintaining their characteristics in serum-free and xeno-free culture conditions.