Cerebellar Development—The Impact of Preterm Birth and Comorbidities

Tam, Emily; Benders, Manon J. N. L.; Heine, Vivi M.

doi:10.1016/b978-0-323-35214-7.00135-9

Cited by 2 publications

(2 citation statements)

References 151 publications

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“…Developmental studies have shown that generation of mid-hindbrain structures involve coordinated activity of different signaling molecules (reviewed by Tam et al, 2017 ). WNT, BMP and fibroblast growth factors (FGFs) in particular set pro-cerebellar patterning of the mid-hindbrain region ( Joyner, 1996 ) and guide formation of the isthmic organizer ( Joyner et al, 2000 ).…”

Section: Introductionmentioning

confidence: 99%

Simplified 3D protocol capable of generating early cortical neuroepithelium

Holmes

Heine

2017

Biology Open

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Here, we report a 3D cerebellar differentiation protocol with quick startup method, defined medium and no special materials or handling requirements. Three fibroblast growth factors (FGF2, 4 and 8) were used for cerebellar patterning and smoothened agonist (SAG) for granule cell development. After 35 days, differentiation products exhibited similar structures and neuronal markers reported in prior ‘organoid’ and ‘spheroid’ protocols. This included cells positive for KIRREL2 (a marker of early cerebellar neuroepithelium) and ZIC1 (a marker for granule cells). Follow-up tests indicated that addition of FGFs, if helpful, was not required to generate observed structures and cell types. This suggests that intrinsic production of patterning factors by aggregates themselves may be adequate for region-specific 3D modeling. This protocol may be used as a quick, easy and cost-efficient method for 3D culture, whether to research development of the early cerebellar neuroepithelium, a base to generate mature cortical structures, or to optimize minimal-factor protocols for other brain regions.

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Section: Introductionmentioning

confidence: 99%

Simplified 3D protocol capable of generating early cortical neuroepithelium

Holmes

Heine

2017

Biology Open

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“…As such, they required a succession of factors introduced at specific times to drive pro-cerebellar patterning and maturation. Chief among these were WNT, bone morphogenetic proteins (BMPs), and fibroblast growth factors (FGFs) with known roles in mid-hindbrain development and formation of the isthmic organizer 5 6 7 . Of course, each additional step and factor means an increase in labor-intensive manipulations and greater expense for the researcher, and so developing simpler protocols capable of achieving equal results is of interest.…”

Section: Introductionmentioning

confidence: 99%

Streamlined 3D Cerebellar Differentiation Protocol with Optional 2D Modification

Holmes

Heine

2017

JoVE

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Reducing the complexity and cost of differentiation protocols is important for researchers. This interest fits with concerns about possible unintended effects that extrinsic patterning factors might introduce into human pluripotent stem cell (hPSC) models of brain development or pathophysiology, such as masking disease phenotype. Here, we present two cerebellar differentiation protocols for hPSCs, designed with simpler startup method, fewer patterning factors, and less material requirements than previous protocols. Recently, we developed culture procedures, which generate free-floating 3-dimensional (3D) products consistent with other brain "organoid" protocols, including morphologies relevant to modeling brain development such as sub/ventricular zone- and rhombic lip-like structures. The second uses an adherent, 2D monolayer procedure to complete differentiation, which is shown capable of generating functional cerebellar neurons, as products are positive for cerebellar-associated markers, and exhibit neuron-like calcium influxes. Together, these protocols offer scientists a choice of options suited to different research purposes, as well as a basic model for testing other types of streamlined neural differentiations.

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Cerebellar Development—The Impact of Preterm Birth and Comorbidities

Cited by 2 publications

References 151 publications

Simplified 3D protocol capable of generating early cortical neuroepithelium

Simplified 3D protocol capable of generating early cortical neuroepithelium

Streamlined 3D Cerebellar Differentiation Protocol with Optional 2D Modification

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