Neurosphere formation enhances the neurogenic differentiation potential and migratory ability of umbilical cord-mesenchymal stromal cells

Cytotherapy. 2016 Feb;18(2):229-41. doi: 10.1016/j.jcyt.2015.10.012.

Abstract

Background aims: The human umbilical cord (UC) is a rich source of mesenchymal stromal cells (MSCs), which have been reported to have multi-lineage potential. The objectives of this study were to investigate the characteristics and capacity of UC-MSC neurosphere formation and whether this event enhances the propensity of UC-MSCs to undergo neural differentiation.

Methods: UC-MSCs were collected by the improved explant method. UC-MSCs and neurosphere-forming UC-MSCs (UC-MSC-neurospheres) were induced to undergo neurogenic differentiation, the latter of which were induced by suspension culturing in the presence of epidermal growth factor and basic fibroblast growth factor. The differentiation and migratory capacities of the individual cultures were then compared on the basis of the expression of neural markers, as measured by immunocytochemistry, immunoblotting and quantitative real-time polymerase chain reaction and transwell assays, respectively.

Results: Both UC-MSCs and UC-MSC-neurospheres were capable of differentiating into neurogenic cells when cultured in neurogenic differentiation medium. However, pre-conditioned UC-MSC-neurospheres exhibited significantly higher expression of neural markers--including microtubule-associated protein (MAP2), MUSASHI1, glial fibrillary acidic protein (GFAP), and NESTIN--compared with those derived from UC-MSCs directly. Moreover, UC-MSC-neurospheres expressed significantly higher levels of the stemness markers NANOG, KLF4 and OCT4 than did UC-MSCs. Migration assays also revealed that both UC-MSCs and UC-MSC-neurospheres actively migrate toward glucose-depleted cells.

Conclusions: Neurogenic differentiation potential probably is greater in UC-MSC-neurospheres than in UC-MSCs. Thus, UC-MSC-neurospheres may serve as a better source of cells for neurogenic regenerative medicine.

Keywords: mesenchymal stromal cell; neural differentiation; neurosphere; umbilical cord.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism
  • Cell Culture Techniques
  • Cell Movement / physiology
  • Epidermal Growth Factor / pharmacology
  • Fibroblast Growth Factor 2 / pharmacology
  • Glial Fibrillary Acidic Protein / biosynthesis
  • Homeodomain Proteins / biosynthesis
  • Humans
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors / biosynthesis
  • Mesenchymal Stem Cells / cytology*
  • Microtubule-Associated Proteins / biosynthesis
  • Nanog Homeobox Protein
  • Nerve Tissue Proteins / biosynthesis
  • Nestin / biosynthesis
  • Neurogenesis / physiology*
  • Octamer Transcription Factor-3 / biosynthesis
  • RNA-Binding Proteins / biosynthesis
  • Real-Time Polymerase Chain Reaction
  • Spheroids, Cellular / cytology*
  • Umbilical Cord / cytology*

Substances

  • Biomarkers
  • Glial Fibrillary Acidic Protein
  • Homeodomain Proteins
  • KLF4 protein, human
  • Kruppel-Like Factor 4
  • Kruppel-Like Transcription Factors
  • MSI1 protein, human
  • Microtubule-Associated Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Nerve Tissue Proteins
  • Nestin
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • RNA-Binding Proteins
  • Fibroblast Growth Factor 2
  • Epidermal Growth Factor