Optimized Induction of Neurospheres from Porcine Mesenchymal Stem Cells

• Published in: The FASEB journal Vol. 32; no. S1; p. 615.6
• Main Authors: Schomberg, Dominic T., Wood, Aleksandar, Singh, Harpreet, Meudt, Jennifer J., Shanmuganayagam, Dhanansayan
• Format: Journal Article
• Language: English
• Published: The Federation of American Societies for Experimental Biology 01-04-2018
• ISSN: 0892-6638; 1530-6860
• DOI: 10.1096/fasebj.2018.32.1_supplement.615.6

Neurospheres are free‐floating clusters of cells that provide researchers a unique method to investigate neural stem cells. Stem cells typically grow as an adherent monolayer on the culture surface, however, when introduced to proper media and growth factors without adherent substrates, these stem cells will form non‐adherent three‐dimensional aggregates called neurospheres. Induction of neurospheres increases multipotency and regenerative capacity of some stem cells, such as adipose‐derived mesenchymal stem cells (adMSCs). Currently, there are no standardized methods for neurosphere induction and evaluation which limits their utility. Studies into cancer, neural development, and regenerative medicine would all benefit from an optimized neurosphere induction protocol. We aimed to establish an optimized protocol for inducing neurospheres from adMSCs in an efficient and reproducible manner.
Primary swine adMSCs were induced to neurospheres with media comprised of varying concentrations of neurobasal medium, B27, and glutamine, epithelial growth factor (EGF) and basic fibroblast growth factor (bFGF) and morphometric parameters of growth (total neurospheres, total cells, cell density and neurosphere diameter) were evaluated daily for eight days. After the eight‐day induction period, the multipotency of neurospheres was determined by assessing presence of neuronal markers nestin, Sox2 and Oct4. We also evaluated for ideal conditions under which neurospheres formed without any signs necrosis at the core. Our optimized neurosphere culture methods produce neural stem cell that best maintain neural‐multipotency and provide a standardized and reproducible protocol for neuronal stem cell propagation.
Support or Funding Information
The research was supported by the Biomedical & Genomic Research Group Discretionary Fund (University of Wisconsin‐Madison), NF North Central, NF Network, and NF Team Foundation.
This is from the Experimental Biology 2018 Meeting. There is no full text article associated with this published in The FASEB Journal.