Transplantation of Bioreactor-Produced Neural Stem Cells into the Rodent Brain

Abstract

The development of new cell replacement strategies using neural stem cells (NSC) may provide an alternative and unlimited cell source for clinical neural transplantation in neurodegenerative diseases such as Parkinson's and Huntington's disease. The clinical application of neural transplantation using NSC will therefore depend upon the availability of clinical grade NSC that are generated in unlimited quantities in a standardized manner. In order to investigate the utility of NSC in clinical neural transplantation, undifferentiated murine NSC were first expanded for an extended period of time in suspension bioreactors containing a serum-free medium. Following expansion in suspension bioreactors, NSC were still able to differentiate in vitro into both astrocytes and neurons after exposure to brain-derived neurotrophic factor (BDNF), suggesting that bioreactor expansion does not alter cell lineage potentiality. Undifferentiated bioreactor-expanded NSC were then transplanted into the rodent striatum. Immunohistochemical examination revealed undifferentiated bioreactor-expanded NSC survived transplantation for up to 8 weeks and expressed the astrocytic immunohistochemical marker glial fibrillary acidic protein (GFAP), suggesting that the host striatal environment influences NSC cell fate upon transplantation. Moreover, no tumor formation was observed within the graft site, indicating that NSC expanded in suspension bioreactors for an extended period of time are a safe source of tissue for transplantation. Future studies should focus on predifferentiating NSC towards specific neuronal phenotypes prior to transplantation in order to restore behavioral function in rodent models of neurodegenerative disease.

Spinal GABAergic Transplants Attenuate Mechanical Allodynia in a Rat Model of Neuropathic Pain

“…Ultimately, differentiation of neural precursor cells into a GABAergic phenotype prior to transplantation may be crucial to avoid the potential deleterious side effects associated with the transplantation of undifferentiated stem cells [107, 108]. Neural stem and precursor cells have typically differentiated into glia after transplantation into nonneurogenic regions of the central nervous system, such as the spinal cord, because of the predominance of glial restrictive cues [109, , , , –114]. The bioreactor‐expanded HNPCs used in our study differentiated into a phenotype immunoreactive for both GABA and TUJ1 in vitro that was maintained even up to 7 weeks after transplantation into the rat spinal cord.…”

Improved expansion of human bone marrow-derived mesenchymal stem cells in microcarrier-based suspension culture

“…shear stresses, hydrodynamic pressures and medium flow patterns, which is known to impact cell characteristics, can be manipulated (Huang et al ., ). We have previously shown that suspension culture bioreactors can be used to expand embryonic stem cells (Cormier et al ., ), neural stem cells (Baghbaderani et al ., , ; Mukhida et al ., ; McLeod et al ., ) and cancer stem cells (Youn et al ., ).…”

Oxygen-Controlled Automated Neural Differentiation of Mouse Embryonic Stem Cells

“…Automated bioreactors have historically allowed the control of mammalian suspension cell culture resulting in repeatable and controlled bioprocesses [13,14]. Although a number of studies have used microcarrier technologies to differentiate ESC in suspension bioreactors [15][16][17][18][19][20][21][22] the vast majority of current differentiation protocols are based on 2D cell cultures.…”