Eliminating the need of serum testing using low serum culture conditions for human bone marrow-derived mesenchymal stromal cell expansion.

BACKGROUND: The conventional expansion of human mesenchymal stromal cells (hMSC) for tissue engineering or (pre-) clinical investigation includes the use of 10% fetal bovine serum (FBS). However, there exists immense lot-to-lot variability in FBS samples and time consuming as well as cost intensive lot pre-testing is essential to guarantee optimal hMSC proliferation and stem cells characteristics maintenance. Furthermore, lot-to-lot variability impedes the long-term consistency of research and comparability between research groups. Therefore, we investigated the use of defined, invariable, non-synthetic FBS in low serum culture conditions for isolation and expansion of hMSC. METHODS: hMSC were isolated from bone marrow in Panserin 401 supplemented with growth factors and 2% MSC-tested or non-tested, defined, invariable, non-synthetic FBS and further cultivated in vitro. The surface marker expression, differentiation capacity as well as cell proliferation and cytotoxicity was analyzed and compared between serum samples. RESULTS: Cells isolated and cultivated with low concentrations of MSC-tested or non-tested FBS demonstrated no differences in surface marker expression or differentiation capacity. Proliferation of hMSC was equal in medium supplemented with either serum with no indication of cell death. CONCLUSIONS: The low serum concentration in Panserin 401 supplemented with growth factors enables the use of defined, invariable, non-synthetic FBS for the isolation and expansion of hMSC. The required hMSC characteristics like surface marker expression and differentiation capacity are maintained. Importantly, no differences in the cell proliferation could be detected. Therefore, using these low-serum culture conditions, the need for lot-to-lot pre-testing of FBS usually needed for optimal hMSC expansion is abolished leading to long-term consistency and comparability of results.

Microstructure and cytocompatibility of collagen matrices for urological tissue engineering.

OBJECTIVES: • To analyse the in vitro cytocompatibility of several engineered collagen-based biomaterials for tissue engineering of the urinary tract. • Tissue-engineered implants for the reconstruction of the urinary tract are of major interest for urological researchers as well as clinicians. Although several materials have been investigated, the ideal replacement has still to be identified. MATERIALS AND METHODS: • Several collagen matrices were tested. • Electron microscopy was used to visualize the microstructure of the tested matrices. • Examination of cell attachment and growth of primary porcine urothelial and smooth muscle cells were performed and cell phenotypes were analysed using immunohistochemical stains. • Urea permeability was investigated using Ussing chamber experiments. RESULTS: • The best cytocompatibility for both urinary tract-specific cell types was obtained with OptiMaix(®) (Matricel GmbH, Herzogenrath, Germany) materials. • Cell-specific phenotypes were maintained during culture as shown by immunohistochemical staining. • Furthermore, simultaneous cultivation of both cell types for 7 and 14 days significantly reduced urea permeability. CONCLUSION: • These results show the potential of OptiMaix materials in tissue engineering approaches of urinary tract tissues.