Design of culture substrates for large-scale expansion of neural stem cells

Konagaya, Shuhei; Kato, Koichi; Nakaji-Hirabayashi, Tadashi; Iwata, Hiroo

doi:10.1016/j.biomaterials.2010.10.008

Cited by 26 publications

(24 citation statements)

References 20 publications

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“…The surface with immobilized LG3 domain of a laminin α1 chain (an α chain contained in laminin‐1) was examined as a culture substrate for the proliferation of NPCs because a chemically defined and xeno‐free cell culture substrate will be required for future clinical applications. LG3 was fused with a His‐tag through recombinant DNA technology and immobilized onto the Ni 2+ ‐bound glass surface through coordination as previously reported (Konagaya et al, ). The amounts of immobilized LG3‐His was determined to be 0.50 ± 0.11 µg/cm 2 .…”

Section: Resultsmentioning

confidence: 99%

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Optimization of surface‐immobilized extracellular matrices for the proliferation of neural progenitor cells derived from induced pluripotent stem cells

Komura

Kato

Konagaya

et al. 2015

Biotech & Bioengineering

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Neural progenitor cells derived from induced pluripotent stem cells have been considered as a potential source for cell-transplantation therapy of central nervous disorders. However, efficient methods to expand neural progenitor cells are further required for their clinical applications. In this study, a protein array was fabricated with nine extracellular matrices and used to screen substrates suitable for the expansion of neural progenitor cells derived from mouse induced pluripotent stem cells. The results showed that neural progenitor cells efficiently proliferated on substrates with immobilized laminin-1, laminin-5, or Matrigel. Based on this result, further attempts were made to develop clinically compliant substrates with immobilized polypeptides that mimic laminin-1, one of the most effective extracellular matrices as identified in the array-based screening. We used here recombinant DNA technology to prepare polypeptide containing the globular domain 3 of laminin-1 and immobilized it onto glass-based substrates. Our results showed that neural progenitor cells selectively proliferated on substrate with the immobilized polypeptide while maintaining their differentiated state.

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Section: Resultsmentioning

confidence: 99%

“…LG3 was fused with a His-tag through recombinant DNA technology and immobilized onto the Ni 2þ -bound glass surface through coordination as previously reported (Konagaya et al, 2011). The amounts of immobilized LG3-His was determined to be 0.50 AE 0.11 mg/cm 2 .…”

Section: Expansion Of Npcs On Lg3-immobilized Substratementioning

confidence: 96%

Optimization of surface‐immobilized extracellular matrices for the proliferation of neural progenitor cells derived from induced pluripotent stem cells

Komura

Kato

Konagaya

et al. 2015

Biotech & Bioengineering

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“…Currently, surface modification with chimeric proteins is being used to develop functional biomaterials worldwide . Previously, a substrate immobilized with chimeric epidermal growth factor (EGF), which acts as a mitogen for neural stem cells and NSPCs, was designed for the uniform expansion of cells expressing only nestin, a marker of neural stem cell . Therefore, we developed a novel culture method, which utilizes an EGF chimeric protein‐anchored substrate to obtain highly pure NSPCs.…”

Section: Introductionmentioning

confidence: 99%

“…13,14 Previously, a substrate immobilized with chimeric epidermal growth factor (EGF), which acts as a mitogen for neural stem cells and NSPCs, was designed for the uniform expansion of cells expressing only nestin, a marker of neural stem cell. [15][16][17][18] Therefore, we developed a novel culture method, which utilizes an EGF chimeric protein-anchored substrate to obtain highly pure NSPCs. We hypothesized that selective differentiation into dopaminergic neurons can be achieved using a system similar to that reported in our previous study.…”

Section: Introductionmentioning

confidence: 99%

Functional surfaces for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons

Nakaji-Hirabayashi

Fujimoto

Yoshikawa

et al. 2019

J Biomedical Materials Res

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Various techniques and systems have been reported for the efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Although a comparatively high percentage of dopaminergic neurons can be obtained using these techniques, the differentiated cells display varied cellular phenotypes such as astrocytes and oligodendrocytes. Generation of highly pure dopaminergic neurons is important for cell‐based therapy and in vitro evaluation of dopaminergic neuron function. In this study, we developed a culture surface anchored with several neurotrophic factors and a neuronal cell‐adhesive protein for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Oligohistidine‐fused brain‐derived neurotrophic factor and glial cell line–derived neurotrophic factor, synthesized using genetic engineering, were co‐immobilized on the surface via metal chelation. To facilitate cell adhesion, a cell‐adhesive chimeric protein derived from laminin (LN‐G) was also immobilized on the surface. Approximately 40% of the cells cultured for 14 days with these protein‐immobilized substrates expressed tyrosine hydroxylase, a marker of dopaminergic neurons, with a three‐fold increase in differentiation efficiency than that reported previously. In addition, the number of tyrosine hydroxylase‐positive cells increased to approximately 80% of the culture after 30 days. These cells secreted dopamine and expressed dopaminergic neuron‐specific genes. Interestingly, cell types (glial cells and oligodendrocytes) other than neuronal cells (immature and mature dopaminergic neurons) were not detected on the protein‐anchored surface. Our results demonstrate that highly pure dopaminergic neurons can be exclusively obtained using the novel substrate without extra purification steps such as cell sorting. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 860–871, 2019.

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“…The third approach is to supply spherical neural masses as a physical method for producing the high yield of dopamine neurons [20,21]. The fourth path is to use biomaterial or combining biomaterial with biological cell growth active factors coated cell culture surfaces [22][23][24]. All of the above are under the ideal natural source conditions, but not applying reprogrammed stem cells.…”

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confidence: 99%

Nonviral Reprogramming Genes Accelerate Formation of Neurons from Murine Embryonic Brain Cells: Synergistic Effect of Brain Derived Neurotrophic Factor Gene Therapy

Liu¹

2016

Transl Med (sunnyvale)

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Design of culture substrates for large-scale expansion of neural stem cells

Cited by 26 publications

References 20 publications

Optimization of surface‐immobilized extracellular matrices for the proliferation of neural progenitor cells derived from induced pluripotent stem cells

Optimization of surface‐immobilized extracellular matrices for the proliferation of neural progenitor cells derived from induced pluripotent stem cells

Functional surfaces for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons

Nonviral Reprogramming Genes Accelerate Formation of Neurons from Murine Embryonic Brain Cells: Synergistic Effect of Brain Derived Neurotrophic Factor Gene Therapy

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