Related Experiment Video
Updated: Jun 22, 2026

Using Primary Neurosphere Cultures to Study Primary Cilia
Published on: April 14, 2017
Properties of secondary and tertiary human enteric nervous system neurospheres
Richard M Lindley1, Daniel B Hawcutt, M Gwen Connell
1Department of Paediatric Surgery, Queens's Medical Centre, Nottingham University Hospitals NHS Trust, Nottingham, UK.
Unlabelled:
Advances in enteric nervous system (ENS) stem cell biology have raised the possibility of treating Hirschsprung's disease with ENS stem/progenitor cell (ENSPC) transplantation. This study aimed to expand ENSPC numbers by the growth and redissociation of neurospheres and assess their differential potential.
Methods:
Human ENS neurospheres were cultured as previously described and redissociated to generate secondary and tertiary neurospheres. Neurospheres were assessed for the presence of neuronal (PGP9.5), glial (S100), and stem cell (p75, nestin markers). The degree of immunofluorescence was quantified using the ImageJ program. Secondary/tertiary neurospheres were transplanted into mouse distal colon grown in tissue culture.
Results:
Secondary/tertiary neurospheres could be generated with exponentially increasing numbers. Tertiary neurospheres showed a significant increase in the proportion of p75 staining but a significant decrease in the proportion of S100 staining. After transplantation, secondary/tertiary neurosphere-derived cells positive for PGP9.5 and S100 could be identified.
Conclusions:
It is possible to exponentially expand neurosphere and therefore ENSPC numbers by repeated dissociation and culture. There is a loss of S100-positive cells in secondary/tertiary neurospheres, but the ENSPCs remain capable of differentiating into neurons and glia when transplanted into an embryonic gut environment.
Related Concept Videos
Peripheral Nervous System: Ganglia and Nerves
Nerves
The nerve is a bundle of axons that serves as the communication highway in the PNS. Each nerve is ensheathed in a protective layer of connective tissue called the epineurium. This outermost layer safeguards the nerve and supports the...
Neurulation
Organization of the Nervous System
The CNS, comprising the brain and spinal cord, houses billions of neurons. The brain is housed in the skull, while the spinal cord is linked to the brain through the foramen magnum of the occipital bone and is surrounded by the protective structure of the vertebral column. It is responsible for processing various...
Nerve Supply of the GI Tract
The enteric nervous system consists of two major plexuses: the myenteric plexus (Auerbach's plexus) and the submucosal plexus (Meissner's plexus). These plexuses are located within the layers of the GI tract...
Functional Divisions of the Nervous System
The sensory division transmits information from sensory receptors in the body to the CNS. It provides the CNS with knowledge about somatic senses (such as tactile, thermal, pain, and proprioceptive sensations)...
Nervous Tissue: Glial Cells
The CNS glial cell includes the astrocytes, the oligodendrocytes, the microglia, and the ependymal cells.
Astrocytes are star-shaped glial cells that interact...

