Efficient animal-serum free 3D cultivation method for adult human neural crest-derived stem cell therapeutics

Due to their broad differentiation potential and their persistence into adulthood, human neural crest-derived stem cells (NCSCs) harbour great potential for autologous cellular therapies, which include the treatment of neurodegenerative diseases and replacement of complex tissues containing various cell types, as in the case of musculoskeletal injuries. The use of serum-free approaches often results in insufficient proliferation of stem cells and foetal calf serum implicates the use of xenogenic medium components. Thus, there is much need for alternative cultivation strategies. In this study we describe for the first time a novel, human blood plasma based semi-solid medium for cultivation of human NCSCs. We cultivated human neural crest-derived inferior turbinate stem cells (ITSCs) within a blood plasma matrix, where they revealed higher proliferation rates compared to a standard serum-free approach. Three-dimensionality of the matrix was investigated using helium ion microscopy. ITSCs grew within the matrix as revealed by laser scanning microscopy. Genetic stability and maintenance of stemness characteristics were assured in 3D cultivated ITSCs, as demonstrated by unchanged expression profile and the capability for self-renewal. ITSCs pre-cultivated in the 3D matrix differentiated efficiently into ectodermal and mesodermal cell types, particularly including osteogenic cell types. Furthermore, ITSCs cultivated as described here could be easily infected with lentiviruses directly in substrate for potential tracing or gene therapeutic approaches. Taken together, the use of human blood plasma as an additive for a completely defined medium points towards a personalisable and autologous cultivation of human neural crest-derived stem cells under clinical grade conditions.

Links between tropical Pacific seasonal, interannual and orbital variability during the Holocene

The El Niño/Southern Oscillation (ENSO) is the leading mode of interannual climate variability. However, it is unclear how ENSO has responded to external forcing, particularly orbitally induced changes in the amplitude of the seasonal cycle during the Holocene. Here we present a reconstruction of seasonal and interannual surface conditions in the tropical Pacific Ocean from a network of high-resolution coral and mollusc records that span discrete intervals of the Holocene. We identify several intervals of reduced variance in the 2 to 7 yr ENSO band that are not in phase with orbital changes in equatorial insolation, with a notable 64% reduction between 5,000 and 3,000 years ago. We compare the reconstructed ENSO variance and seasonal cycle with that simulated by nine climate models that include orbital forcing, and find that the models do not capture the timing or amplitude of ENSO variability, nor the mid-Holocene increase in seasonality seen in the observations; moreover, a simulated inverse relationship between the amplitude of the seasonal cycle and ENSO-related variance in sea surface temperatures is not found in our reconstructions. We conclude that the tropical Pacific climate is highly variable and subject to millennial scale quiescent periods. These periods harbour no simple link to orbital forcing, and are not adequately simulated by the current generation of models.