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


OBJECTIVE
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.


CONCLUSIONS
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.
Gu
Haigang
H
Division of Materials Technology, School of Materials Science & Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore. lptan@ntu.edu.sg
Yue
Zhilian
Z
Leong
Wen Shing
WS
Nugraha
Bramasta
B
Tan
Lay Poh
LP
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002482:Cellulose / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000494:pharmacology*; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000187:drug effects* / Q000648:ultrastructure; D016062:Porosity / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D054457:Tissue Scaffolds
D051792:Basic Helix-Loop-Helix Proteins; D002482:Cellulose; D005904:Glial Fibrillary Acidic Protein; D020100:Hydrogels; D007381:Intermediate Filament Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D012333:RNA, Messenger; C489788:MAP2 protein, human; C577484:NES protein, human; C093564:Neurogenic differentiation factor 1