Bioprocessing strategies to enhance the challenging isolation of neuro-regenerative cells from olfactory mucosa

Scientific Reports, 2018 · DOI: 10.1038/s41598-018-32748-w · Published: October 5, 2018

Neurology Genetics Regenerative Medicine & Stem Cells

Simple Explanation

Olfactory ensheathing cells (OECs) are being explored as a potential cell therapy for spinal cord injury repair due to their ability to support nerve regeneration. These cells are located in the olfactory bulb in the brain and the olfactory mucosa within the nasal cavity. This study aims to optimize the isolation and culture methods for OECs derived from the olfactory mucosa. The research investigates the effects of different bioprocess modifications on the resulting cell population from rat olfactory mucosal tissue, focusing on the expression of key OEC markers. The key bioprocess conditions studied include cell culture substrate, serum concentration, oxygen tension, enrichment with neurotrophic factor-3 (NT-3), and differential adhesion. Following the optimization, the ability of the isolated OECs to support and promote neuronal growth in 2D neuron co-culture in vitro was assessed.

Study Duration

14 days

Participants

Adult Sprague-Dawley female rats (200–250 g)

Evidence Level

Not specified

Key Findings

1
A 24-hour differential adhesion step significantly increased the expression of p75NTR, an OEC marker, to 73 ± 5% and 46 ± 18% on PDL and laminin matrices, respectively.
2
The introduction of neurotrophic factor NT-3 and a decrease in serum concentration to 2% FBS resulted in enrichment of OECs, with p75NTR expression reaching nearly 100% and the fibroblast marker Thy1.1 decreasing to zero.
3
Culturing OECs at physiologically relevant oxygen tension (2–8%) had a negative impact on p75NTR expression and overall cell survival. Moreover, co-culture of OECs with NG108-15 neurons resulted in more neuronal growth and potential migration at atmospheric oxygen.

Research Summary

This study explored methods to enhance the recovery of cells expressing OEC marker p75NTR from rat mucosa. Key bioprocessing fundamentals were identified that will underpin future development of OEC-based cell therapies for potential use in spinal cord injury repair. The optimum culturing conditions for primary rat OECs have been identified as a concentration of 2% FBS and a concentration of 50 ng/ml NT-3 on laminin coating.

Practical Implications

Optimized OEC Isolation

The identified bioprocessing strategies enhance the purity and yield of OECs from olfactory mucosa, making them more suitable for cell therapies.

Enhanced Neuronal Growth

OECs cultured under optimized conditions promote neuronal growth and migration, suggesting their potential in spinal cord injury repair.

Standardized Method Development

The research contributes to the development of a standardized method for OEC isolation, leading to more reproducible outcomes in cell therapy applications.

Study Limitations

1
Inconsistencies in yields and purities obtained across independent experiments.
2
Source of variability in OEC isolation is presently not known.
3
Further work needed to create standardized and consistent isolation methods.

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