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

Abstract

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 insuffi cient 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 fi rst 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 profi le and the capability for self-renewal. ITSCs pre-cultivated in the 3D matrix differentiated effi ciently 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 defi ned medium points towards a personalisable and autologous cultivation of human neural crest-derived stem cells under clinical grade conditions.

Translational Research: Palatal-derived Ecto-mesenchymal Stem Cells from Human Palate: A New Hope for Alveolar Bone and Cranio-Facial Bone Reconstruction

“…Although it is clear that cells residing in the subepithelial soft tissue of the palate can achieve bone regeneration, this population is heterogenous (Harris, 1999), but it is not known which subpopulations are capable of achieving regeneration. Indeed, cells derived from palate were found to have specific properties, such as increased proliferation rates, and representative of a regenerative phenotype as shown by research group (Greiner et al 2011, Widera et al 2009). …”

Comparison of Different Sources of Mesenchymal Stem Cells: Palatal versus Lipoaspirated Adipose Tissue

“…For example, one study demonstrated that palatal tissue-derived stem cells in the subepithelial soft tissue of the palate could be used in bone regeneration [Harris, 1999]. Other research showed that stem cells derived from palatal tissue had specific properties, such as increased proliferation rates and a regenerative phenotype [Widera et al, 2009;Greiner et al, 2011]. Roman et al [2013] were the first to identify palatal tissue-derived MSCs and named them progenitor-like cells.…”

Label-free nonlinear optical microscopy detects early markers for osteogenic differentiation of human stem cells

“…At a confluency of approximately 80%, osteogenic induction medium was applied containing DMEM, 10% FCS, 100 nM dexamethasone (Sigma-Aldrich, St. Louis, USA), 10 mM β -glycerophosphate (Sigma-Aldrich, St. Louis, USA) and 0.05 mM L-ascorbic acid-2-phosphate (Sigma-Aldrich, St. Louis, USA) at 37 °C and 5% CO 2 in a humidified incubator as described by Greiner et al 21. Control cells were cultured in DMEM supplemented with 10% FCS.…”

“…Inferior turbinate stem cells (ITSCs) are routinely isolated during minimally-invasive surgery20 and can easily be expanded in vitro 21 under clinical-grade conditions22. In vitro , ITSCs revealed an extraordinarily broad differentiation potential into ectodermal as well as mesodermal cell types212324.…”