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Table 2 Summary of protocols to obtain neural trans-differentiation of hADSCs

From: Advances, challenges and future directions for stem cell therapy in amyotrophic lateral sclerosis

Culture method Differentiation protocol Neuronal/glial marker expressed Comments Reference
Adherent method 24 h pre-incubation in DMEM/FCS 20% followed by 24 h incubation in DMEM containing BHA, valproic acid, forskolin, hydrocortisone and insulin. Nestin (developing nervous system cells) NeuN (neuronal nuclei) GFAP (mature astrocytes) I-NFM (Neurons) Cells started to lose their neuronal morphology after 4–5 days and all died in 14 days. [88]
Adherent method 14 days incubation in DMEM containing insulin, indomethacin and IBMX Vimentin (Schwann cells) Trk-A (central nervous system) NSE (early neuronal progenitors) Low neuronal marker expression also in undifferentiated ADSCs. 25% differentiation rate obtained. Cells not able to generate action potentials. [90]
Adherent method with or without human Schwann cells 24 h pre-incubation in DMEM/FBS 20% and β-mercaptoethanol followed by 8-16 h incubation in DMEM containing DMSO, β-mercaptoethanol and BHA GFPA S-100 (Astrocytes and Schwann cells) NeuN Nestin Gal-C (Oligodendrocyte) Cell morphology turned back to fibroblast shape after 72 h in normal basal medium. Co-culture with human irradiated Schwann cells enhanced survival (12 days) and expression of myelin proteins. [91]
Adherent method 7 days incubation in DMEM containing bFGF followed by 7 days incubation in DMEM containing forskolin GFPA I-NFM Tuj −1 (neuron specific β-III tubulin) Nestin SNAP-25 (synaptic marker) CNPase (Oligodendrocyte) Inward and outward ion current in patch-clamp experiment. High mRNA expression of ion channels. Low differentiation marker expression also in undifferentiated ADSCs. [92]
Neurosphere method followed by maturation on poly-D-lysine 8 days pre-incubation in DMEM containing β-mercaptoethanol and b-FGF followed by 7 days incubation in neural basal medium (N2B27) and further 7 days in N2B27 medium containing EGF and bGFG. Final maturation achieved from neurosphere dissociated progenitors after 14 day culture in N2B27 media containing retinoic acid and BDNF MAP2 (mature neurons) Nestin Sox1(neural tube development) Pax6 (human neuroepithelium) Vimentin Inward and outward ion currents in patch-clamp experiment. High mRNA expression of ion channels [95]
  1. DMEM Dulbecco’s modified eagle medium, FCS foetal calf serum, BHA butylated hydroxyanisole, GFAP glial fibrillary acid protein, I-NFM intermediate neurofilament, IBMX isobutylmethylxantine, Trk-A tropomyosin receptor kinase A, NSE neuron specific enolase, DMSO dimethyl sulfoxide, S-100 S protein 100, Gal-C galactosylceramidase, bFGF basic fibroblast growth factor, CNPase 2,3-Cyclic-nucleotide 3-phosphodiesterase, EGF epidermal growth factor, BDNF brain derived neurotrophic factor, MAP2 microtubule-associated protein 2