3-Dimensional, Dynamic Cultivations of Human Umbilical Cord-Derived Cells

Abstract

The regeneration of critical sized bone defects remains challenging and for an engineering of biologically functional bone substitutes it is of crucial importance to establish reproducible methods for the systematic differentiation of easily accessible human stem cells in a controlled environment. In this work, the influence of a 3-dimensional macroporous ceramic scaffold (Sponceram (R)) on the differentiation of human umbilical cord-derived cells (hUCs) was investigated under dynamic conditions using a disposable rotating bed bioreactor (Z (R) RPD). Cells were cultivated on Sponceram (R)-discs for 35 days in the Z (R) RPD in standard or osteogenic differentiation medium. The results reveal that cultivation of hUCs in the Z (R) RPD bioreactor are feasible for at least up to 35 days without loss of cell viability as proven by a growing glucose consumption. The cells establish their own extracellular matrix (ECM) in massive amounts and seem to differentiate into the osteogenic lineage even without addition of differentiating factors as shown by histological stainings, flowcytometric analysis and a RT-PCR of common osteogenic mRNAs. Therefore the disposable Z (R) RPD bioreactor, together with the Sponceram (R) carriers, provides an efficient tool for the progeny and osteogenic differentiation of hUCs.