Control of in vitro neural differentiation of mesenchymal stem cells in 3D macroporous, cellulosic hydrogels

Regen Med. 2010 Mar;5(2):245-53. doi: 10.2217/rme.09.89.

Abstract

Background: Mesenchymal stem cells (MSCs) are multipotent cells that can be induced to differentiate into multiple cell lineages, including neural cells. They are a good cell source for neural tissue-engineering applications. Cultivation of human (h)MSCs in 3D scaffolds is an effective means for the development of novel neural tissue-engineered constructs, and may serve as a promising strategy in the treatment of nerve injury.

Aim: This study presents the in vitro growth and neural differentiation of hMSCs in 3D macroporous, cellulosic hydrogels.

Results: The number of hMSCs cultivated in the 3D scaffolds increased by more than 14-fold after 7 days. After 2 days induction, most of the hMSCs in the 3D scaffolds were positive for nestin, a marker of neural stem cells. After 7 days induction, most of the hMSCs in the 3D scaffolds showed glial fibrillary acidic protein, tubulin or neurofilament M-positive reaction and a few hMSCs were positive for nestin. After 14 days induction, hMSCs in the 3D scaffolds could completely differentiate into neurons and glial cells. The neural differentiation of hMSCs in the 3D scaffolds was further demonstrated by real-time PCR.

Conclusion: These results show that the 3D macroporous cellulosic hydrogel could be an appropriate substrate for neural differentiation of hMSCs and its possible applications in neural tissue engineering are discussed.

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cattle
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Cell Shape / drug effects
  • Cellulose / pharmacology*
  • Gene Expression Regulation / drug effects
  • Glial Fibrillary Acidic Protein / genetics
  • Glial Fibrillary Acidic Protein / metabolism
  • Humans
  • Hydrogels / pharmacology*
  • Intermediate Filament Proteins / genetics
  • Intermediate Filament Proteins / metabolism
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects*
  • Mesenchymal Stem Cells / ultrastructure
  • Microscopy, Confocal
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Nestin
  • Neurons / cytology*
  • Neurons / drug effects*
  • Neurons / ultrastructure
  • Porosity / drug effects
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Time Factors
  • Tissue Scaffolds

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Glial Fibrillary Acidic Protein
  • Hydrogels
  • Intermediate Filament Proteins
  • MAP2 protein, human
  • Microtubule-Associated Proteins
  • NES protein, human
  • Nerve Tissue Proteins
  • Nestin
  • RNA, Messenger
  • NeuroD protein
  • Cellulose