journal article·2018-07Open access

An integrated biomanufacturing platform for the large-scale expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells

Gayathri Srinivasan, Daylin Morgan, Divya Varun, Nicholas Brookhouser, David A. Brafman

Acta Biomaterialia· Volume 74

Abstract

Human pluripotent stem cell derived neural progenitor cells (hNPCs) have the unique properties of long-term in vitro expansion as well as differentiation into the various neurons and supporting cell types of the central nervous system (CNS). Because of these characteristics, hNPCs have tremendous potential in the modeling and treatment of various CNS diseases and disorders. However, expansion and neuronal differentiation of hNPCs in quantities necessary for these applications is not possible with current two dimensional (2-D) approaches. Here, we used a fully defined peptide substrate as the basis for a microcarrier (MC)-based suspension culture system. Several independently derived hNPC lines were cultured on MCs for multiple passages as well as efficiently differentiated to neurons. Finally, this MC-based system was used in conjunction with a low shear rotating wall vessel (RWV) bioreactor for the integrated, large-scale expansion and neuronal differentiation of hNPCs. Overall, this fully defined and scalable biomanufacturing system will facilitate the generation of hNPCs and their neuronal derivatives in quantities necessary for basic and translational applications. Statement of significance In this work, we developed a microcarrier (MC)-based culture system that allows for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells (hNPCs) under defined conditions. In turn, this MC approach was implemented in a rotating wall vessel (RWV) bioreactor for the large-scale expansion and neuronal differentiation of hNPCs. This work is of significance as it overcomes current limitations of conventional two dimensional (2-D) culture systems to enable the generation of hNPCs and their neuronal derivatives in quantities required for downstream applications in disease modeling, drug screening, and regenerative medicine.
Source: europepmc · Quality: candidate unverified · Source record ↗

Research context

OpenAlex topics: Pluripotent Stem Cells Research · Neurogenesis and neuroplasticity mechanisms · Neuroscience and Neural Engineering

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

References (89)

Show source-provided references
1. Wnt/Yes-associated protein interactions during neural tissue patterning of human induced pluripotent stem cells
2. Patient-derived iPSCs show premature neural differentiation and neuron type-specific phenotypes relevant to neurodevelopment
3. Endogenous WNT signaling regulates hPSC-derived neural progenitor cell heterogeneity and specifies their regional identity10.1016/j.stemcr.2014.10.004 ↗
4. Neural subtype specification from human pluripotent stem cells10.1016/j.stem.2016.10.015 ↗
5. Advances in stem cell research- a ray of hope in better diagnosis and prognosis in neurodegenerative diseases10.3389/fmolb.2016.00072 ↗
6. Potential of GABA-ergic cell therapy for schizophrenia, neuropathic pain, and Alzheimer's and Parkinson's diseases10.1016/j.brainres.2015.09.019 ↗
7. Application of human induced pluripotent stem cells for modeling and treating neurodegenerative diseases10.1016/j.nbt.2014.05.001 ↗
8. Induced pluripotent stem cells for disease modeling and drug discovery in neurodegenerative diseases10.1007/s12035-014-8867-6 ↗
9. Process engineering of human pluripotent stem cells for clinical application10.1016/j.tibtech.2012.03.003 ↗
10. Neural differentiation from pluripotent stem cells: the role of natural and synthetic extracellular matrix10.4252/wjsc.v6.i1.11 ↗
11. Efficient and fast differentiation of human neural stem cells from human embryonic stem cells for cell therapy10.1155/2017/9405204 ↗
12. Cell-surface marker signatures for the isolation of neural stem cells, glia and neurons derived from human pluripotent stem cells10.1371/journal.pone.0017540 ↗
13. Materials for stem cell factories of the future10.1038/nmat3972 ↗
14. A small molecule screen in stem-cell-derived motor neurons identifies a kinase inhibitor as a candidate therapeutic for ALS10.1016/j.stem.2013.04.003 ↗
15. High-content screening in hPSC-neural progenitors identifies drug candidates that inhibit zika virus infection in fetal-like organoids and adult brain10.1016/j.stem.2017.06.017 ↗
16. Human pluripotent stem cell-derived cortical neurons for high throughput medication screening in autism: a proof of concept study in SHANK3 haploinsufficiency syndrome10.1016/j.ebiom.2016.05.032 ↗
17. Utilization of iPSC-derived human neurons for high-throughput drug-induced peripheral neuropathy screening10.1016/j.tiv.2017.08.014 ↗
18. Large-scale production of human pluripotent stem cell derived cardiomyocytes10.1016/j.addr.2015.11.016 ↗
19. Scalable stirred suspension culture for the generation of billions of human induced pluripotent stem cells using single-use bioreactors
20. Neural stem cells: ready for therapeutic applications?10.1186/2052-8426-2-31 ↗
21. Human neural precursor cells promote neurologic recovery in a viral model of multiple sclerosis10.1016/j.stemcr.2014.04.005 ↗
22. Human induced pluripotent stem cells are a novel source of neural progenitor cells (iNPCs) that migrate and integrate in the rodent spinal cord10.1002/cne.23578 ↗
23. Neural and glial progenitor transplantation as a neuroprotective strategy for Amyotrophic Lateral Sclerosis (ALS)10.1016/j.brainres.2015.06.035 ↗
24. Long-distance growth and connectivity of neural stem cells after severe spinal cord injury10.1016/j.cell.2012.08.020 ↗
25. Neural stem cell or human induced pluripotent stem cell-derived GABA-ergic progenitor cell grafting in an animal model of chronic temporal lobe epilepsy10.1002/cpsc.9 ↗
26. Generation of high-purity human ventral midbrain dopaminergic progenitors for in vitro maturation and intracerebral transplantation10.1038/nprot.2017.078 ↗
27. ESC-Derived basal forebrain cholinergic neurons ameliorate the cognitive symptoms associated with alzheimer's disease in mouse models10.1016/j.stemcr.2015.09.010 ↗
28. A compendium of preparation and application of stem cells in parkinson's disease: current status and future prospects10.3389/fnagi.2016.00117 ↗
29. Transplantation of feeder-free human induced pluripotent stem cell-derived cortical neuron progenitors in adult male Wistar rats with focal brain ischemia
30. Patient-derived hiPSC neurons with heterozygous CNTNAP2 deletions display altered neuronal gene expression and network activity10.1038/s41537-017-0033-5 ↗
31. Altered differentiation potential of Gaucher's disease iPSC neuronal progenitors due to Wnt/beta-catenin downregulation10.1016/j.stemcr.2017.10.029 ↗
32. iPS-derived neural progenitor cells from PPMS patients reveal defect in myelin injury response10.1016/j.expneurol.2016.11.012 ↗
33. Zika virus infection induces mitosis abnormalities and apoptotic cell death of human neural progenitor cells10.1038/srep39775 ↗
34. Highly expandable human iPS cell-derived neural progenitor cells (NPC) and neurons for central nervous system disease modeling and high-throughput screening
35. Recent Zika virus isolates induce premature differentiation of neural progenitors in human brain organoids10.1016/j.stem.2016.12.005 ↗
36. Induced pluripotent stem cells in Alzheimer's disease: applications for disease modeling and cell-replacement therapy10.1186/s13024-016-0106-3 ↗
37. Modeling ALS with motor neurons derived from human induced pluripotent stem cells10.1038/nn.4273 ↗
38. The progress of induced pluripotent stem cells as models of parkinson's disease10.1155/2016/4126214 ↗
39. A robust vitronectin-derived peptide for the scalable long-term expansion and neuronal differentiation of human pluripotent stem cell10.1016/j.actbio.2016.10.037 ↗
40. Novel quantitative biosystem for modeling physiological fluid shear stress on cells10.1128/AEM.02428-06 ↗
41. Optimized suspension culture: the rotating-wall vessel10.1152/ajprenal.2001.281.1.F12 ↗
42. Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro10.1038/74447 ↗
43. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA10.1016/j.stem.2010.08.012 ↗
44. Generation and characterization of human induced pluripotent stem cell (hiPSC) lines from an Alzheimer's disease (ASUi003-A) and non-demented control (ASUi004-A) patient homozygous for the Apolipoprotein e4 (APOE4) risk variant10.1016/j.scr.2017.07.003 ↗
45. APOE epsilon4/epsilon4 diminishes neurotrophic function of human iPSC-derived astrocytes10.1093/hmg/ddx155 ↗
46. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method10.1006/meth.2001.1262 ↗
47. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome10.1186/gb-2009-10-3-r25 ↗
48. HISAT: a fast spliced aligner with low memory requirements10.1038/nmeth.3317 ↗
49. RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome10.1186/1471-2105-12-323 ↗
50. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation10.1038/nbt.1621 ↗
51. The significance of digital gene expression profiles10.1101/gr.7.10.986 ↗
52. NeuN As a neuronal nuclear antigen and neuron differentiation marker10.32607/20758251-2015-7-2-42-47 ↗
53. Cell biology. Deconstructing dimensionality10.1126/science.1233814 ↗
54. Taking cell-matrix adhesions to the third dimension10.1126/science.1064829 ↗
55. Cell interactions with three-dimensional matrices10.1016/S0955-0674(02)00364-2 ↗
56. Differentiation patterns of embryonic stem cells in two- versus three-dimensional culture10.1159/000346166 ↗
57. 3D culture models of Alzheimer's disease: a road map to a “cure-in-a-dish”10.1186/s13024-016-0139-7 ↗
58. A three-dimensional human neural cell culture model of Alzheimer's disease10.1038/nature13800 ↗
59. Application of human mesenchymal and pluripotent stem cell microcarrier cultures in cellular therapy: achievements and future direction10.1016/j.biotechadv.2013.03.006 ↗
60. Microcarrier culture systems
61. Engineering stem cell fate with biochemical and biomechanical properties of microcarriers10.1002/btpr.1825 ↗
62. Critical microcarrier properties affecting the expansion of undifferentiated human embryonic stem cells10.1016/j.scr.2011.04.007 ↗
63. Dynamic suspension culture for scalable expansion of undifferentiated human pluripotent stem cells10.1038/nprot.2011.325 ↗
64. The multiparametric effects of hydrodynamic environments on stem cell culture10.1089/ten.teb.2011.0040 ↗
65. A review of bioreactor protocols for human neural precursor cell expansion in preparation for clinical trials10.2174/157488811796575378 ↗
66. Progress and challenges in large-scale expansion of human pluripotent stem cells10.1016/j.procbio.2016.09.032 ↗
67. Expansion of mammalian neural stem cells in bioreactors: effect of power input and medium viscosity10.1016/S0165-3806(01)00328-5 ↗
68. Transplantation of bioreactor-produced neural stem cells into the rodent brain10.3727/000000006783464426 ↗
69. Novel bioreactors for the culture and expansion of aggregative neural stem cells10.1007/s00449-007-0174-4 ↗
70. Neural stem cell systems: physiological players or in vitro entities?10.1038/nrn2761 ↗
71. Markers of pluripotency and differentiation in human neural precursor cells derived from embryonic stem cells and CNS tissue10.3727/096368910X527266 ↗
72. Microcarrier-expanded neural progenitor cells can survive, differentiate, and innervate host neurons better when transplanted as aggregates10.3727/096368915X690378 ↗
73. Microcarrier suspension cultures for high-density expansion and differentiation of human pluripotent stem cells to neural progenitor cells10.1089/ten.tec.2012.0146 ↗
74. Selective neuronal vulnerability in neurodegenerative diseases: from stressor thresholds to degeneration10.1016/j.neuron.2011.06.031 ↗
75. Cell-type dependent alzheimer's disease phenotypes: probing the biology of selective neuronal vulnerability10.1016/j.stemcr.2017.10.015 ↗
76. Controlling the regional identity of hPSC-derived neurons to uncover neuronal subtype specificity of neurological disease phenotypes10.1016/j.stemcr.2015.10.005 ↗
77. Astrocytes in alzheimer's disease10.1016/j.nurt.2010.05.017 ↗
78. Astrocytes in physiological aging and Alzheimer's disease10.1016/j.neuroscience.2015.01.007 ↗
79. Astrocytes in neurodegenerative disease10.1101/cshperspect.a020628 ↗
80. Specification of neuronal and glial subtypes from human pluripotent stem cells10.1007/s00018-011-0770-y ↗
81. Capturing complex 3D tissue physiology in vitro10.1038/nrm1858 ↗
82. 3D differentiation of neural stem cells in macroporous photopolymerizable hydrogel scaffolds10.1371/journal.pone.0048824 ↗
83. Cerebral organoids model human brain development and microcephaly10.1038/nature12517 ↗
84. Alzheimer's in 3D culture: challenges and perspectives10.1002/bies.201500063 ↗
85. Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture10.1038/nmeth.3415 ↗
86. Modeling mouse and human development using organoid cultures10.1242/dev.118570 ↗
87. The use of brain organoids to investigate neural development and disease10.1038/nrn.2017.107 ↗
88. Dishing out mini-brains: Current progress and future prospects in brain organoid research10.1016/j.ydbio.2016.06.037 ↗
89. Quantitation of apoptosis and necrosis by Annexin V Binding, propidium iodide uptake, and flow cytometry10.1101/pdb.prot087288 ↗