xeno-free culture condition for human bone marrow and umbilical cord matrix-derived mesenchymal stem/stromal cells using human umbilical cord blood serum

Background: fetal bovine serum (fbs) is widely used in cell culture laboratories,risk of zoonotic infections and allergic side effects create obstacles for its use inclinical trials. therefore, an alternative supplement with proper inherent growthpromotingactivities is demanded.objective: to find fbs substitute, we tested human umbilical cord blood serum(hucs) for proliferation of human umbilical cord matrix derived mesenchymal stemcells (huc-mscs) and human bone marrow-derived mesenchymal cells (hbmmscs).materials and methods: umbilical cord blood of healthy neonates, delivered bycaesarian section, was collected and the serum was separated. huc-mscs andhbm-mscs were isolated and characterized by assessment of cell surface antigensby flow cytometry, alkaline phosphatase activity and osteogenic/adipogenicdifferentiation potential. the cells were then cultured in iscove's modifieddulbecco's medium (imdm) by conventional methods in three preparations: 1- withhucs, 2- with fbs, and 3- without serum supplements. cell proliferation wasmeasured using wst-1 assay, and cell viability was assessed by trypan bluestaining.results: the cells cultured in hucs and fbs exhibited similar morphology andmesenchymal stem cells properties. wst-1 proliferation assay data showed nosignificant difference between the proliferation rate of either cells following hucsand fbs supplementation. trypan blue exclusion dye test also revealed nosignificant difference for viability between hucs and fbs groups. a significantdifference was detected between the proliferation rate of stem cells cultured inserum-supplemented medium compared with serum-free medium.conclusion: our results indicate that human umbilical cord serum can effectivelysupport proliferation of hbm-mscs and huc-mscs in vitro and can be used as anappropriate substitute for fbs, especially in clinical studies.