High-Yield Mucosal Olfactory Ensheathing Cells Restore Loss of Function in Rat Dorsal Root Injury

In a previous study, we reported that no axons were crossing from the severed dorsal roots to the spinal cord using the rat dorsal rhizotomy paradigm. The injury caused ipsilateral deficits of forepaw function. An attempt to restore the function by transplanting cells containing 5% olfactory ensheathing cells (OECs) cultured from the olfactory mucosa did not succeed. However, obtaining OECs from the olfactory mucosa has an advantage for clinical application. In the present study, we used the same rhizotomy paradigm, but rats with an injury received cells from a modified mucosal culture containing around 20% OECs mixed in collagen. The forelimb proprioception assessment showed that 80% of the rats receiving the transplants had functional improvement over six weeks of the study. The adhesive removal test showed that the time taken for the rats to notice the adhesive label and remove it almost returned to the normal level after receiving the transplants. Transplanted cells were identified with the expression of green fluorescent protein (ZsGreen). Some regeneration fibres immunostained for neurofilament (NF) or traced by biotinylated dextran amine (BDA) in the injury area were associated with the transplanted cells. The evidence in this study improves the prospect of clinical application using OECs from the olfactory mucosa to treat CNS injuries.

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Olfactory ensheathing cells from the nose: Clinical application in human spinal cord injuries

Experimental Neurology ◽

10.1016/j.expneurol.2010.08.025 ◽

2011 ◽

Vol 229 (1) ◽

pp. 174-180 ◽

Cited By ~ 81

Author(s):

Alan Mackay-Sim ◽

James A. St John

Keyword(s):

Spinal Cord ◽

Clinical Application ◽

Spinal Cord Injuries ◽

Olfactory Ensheathing Cells ◽

Human Spinal Cord ◽

Ensheathing Cells

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Could Cells from Your Nose Fix Your Heart? Transplantation of Olfactory Stem Cells in a Rat Model of Cardiac Infarction

The Scientific World JOURNAL ◽

10.1100/tsw.2010.40 ◽

2010 ◽

Vol 10 ◽

pp. 422-433 ◽

Cited By ~ 11

Author(s):

Cameron McDonald ◽

Alan Mackay-Sim ◽

Denis Crane ◽

Wayne Murrell

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Fluorescent Protein ◽

Genetically Engineered ◽

Olfactory Mucosa ◽

Cell Transplant ◽

Adult Rat ◽

Cardiac Ventricle ◽

Green Fluorescent

This study examines the hypothesis that multipotent olfactory mucosal stem cells could provide a basis for the development of autologous cell transplant therapy for the treatment of heart attack. In humans, these cells are easily obtained by simple biopsy. Neural stem cells from the olfactory mucosa are multipotent, with the capacity to differentiate into developmental fates other than neurons and glia, with evidence of cardiomyocyte differentiationin vitroand after transplantation into the chick embryo. Olfactory stem cells were grown from rat olfactory mucosa. These cells are propagated as neurosphere cultures, similar to other neural stem cells. Olfactory neurospheres were grownin vitro, dissociated into single cell suspensions, and transplanted into the infarcted hearts of congeneic rats. Transplanted cells were genetically engineered to express green fluorescent protein (GFP) in order to allow them to be identified after transplantation. Functional assessment was attempted using echocardiography in three groups of rats: control, unoperated; infarct only; infarcted and transplanted. Transplantation of neurosphere-derived cells from adult rat olfactory mucosa appeared to restore heart rate with other trends towards improvement in other measures of ventricular function indicated. Importantly, donor-derived cells engrafted in the transplanted cardiac ventricle and expressed cardiac contractile proteins.

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Gene and protein expression profiles of olfactory ensheathing cells from olfactory bulb versus olfactory mucosa

Neural Regeneration Research ◽

10.4103/1673-5374.317986 ◽

2022 ◽

Vol 17 (2) ◽

pp. 440

Author(s):

Li-Bin Wang ◽

He-Chun Xia ◽

Yuan-Xiang Lan ◽

Ping Yang ◽

Zhong Zeng ◽

...

Keyword(s):

Olfactory Bulb ◽

Protein Expression ◽

Expression Profiles ◽

Olfactory Mucosa ◽

Olfactory Ensheathing Cells ◽

Gene And Protein Expression ◽

Ensheathing Cells ◽

Protein Expression Profiles

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OBTAINING OF CELLULAR PREPARATIONS OF RAT AND HUMAN OLFACTORY MUCOSA AND THEIR INFLUENCE ON THE SIZE OF MODELED SPINAL CORD CYSTALS

BIOTECHNOLOGY: STATE OF THE ART AND PERSPECTIVES ◽

10.37747/2312-640x-2021-19-75-77 ◽

2021 ◽

Vol 1 (19) ◽

pp. 75-77

Author(s):

O.V. Stepanova ◽

E.K. Karsuntseva ◽

G.A. Fursa ◽

A.V. Chadin ◽

M.P. Valikhov ◽

...

Keyword(s):

Spinal Cord ◽

Progenitor Cells ◽

Olfactory Mucosa ◽

Olfactory Ensheathing Cells ◽

Complete Disappearance ◽

Ensheathing Cells ◽

Neural Stem Progenitor Cells ◽

Human Olfactory Mucosa ◽

Enriched Cultures

Enriched cultures of olfactory ensheathing cells and neural stem/progenitor cells were obtained according to our developed protocols from the olfactory mucosa of rat and human. It has been shown that only transplantation of human and rat olfactory ensheathing cells leads to a significant decrease in the size of cysts, as well as their complete disappearance in some animals.

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A conditional mouse line for lineage tracing of Sox9 loss-of-function cells using enhanced green fluorescent protein

Biotechnology Letters ◽

10.1007/s10529-013-1303-6 ◽

2013 ◽

Vol 35 (12) ◽

pp. 1991-1996 ◽

Cited By ~ 3

Author(s):

Sumantra Chatterjee ◽

Petra Kraus ◽

V. Sivakamasundari ◽

Xing Xing ◽

Sook Peng Yap ◽

...

Keyword(s):

Green Fluorescent Protein ◽

Enhanced Green Fluorescent Protein ◽

Fluorescent Protein ◽

Lineage Tracing ◽

Mouse Line ◽

Loss Of Function ◽

Green Fluorescent

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In vitro Maintenance of Olfactory Mucosa: with Enriched Olfactory Ensheathing Cells

Journal of Stem Cell Research & Therapy ◽

10.4172/2157-7633.1000132 ◽

2013 ◽

Vol 03 (01) ◽

Cited By ~ 4

Author(s):

Neetu Singh ◽

Saroj Chooramani Gopal ◽

Rajeshwar Nath Srivastava ◽

Tulika Chandra ◽

Satya Prakash Agarwal

Keyword(s):

Olfactory Mucosa ◽

Olfactory Ensheathing Cells ◽

Ensheathing Cells ◽

In Vitro Maintenance

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Gli3 regulates vomeronasal neurogenesis, olfactory ensheathing cell formation and GnRH-1 neuronal migration

10.1101/643155 ◽

2019 ◽

Author(s):

Ed Zandro M. Taroc ◽

Ankana Naik ◽

Jennifer M. Lin ◽

Nicolas B. Peterson ◽

David L. Keefe ◽

...

Keyword(s):

Transcription Factor ◽

Nasal Mucosa ◽

Neuronal Migration ◽

Neuronal Development ◽

Olfactory Ensheathing Cells ◽

Loss Of Function ◽

Sequencing Data ◽

Whole Exome ◽

Ensheathing Cells ◽

Whole Exome Sequencing Data

AbstractDuring mammalian development, gonadotropin-releasing-hormone-1 neurons (GnRH-1ns) migrate from the developing vomeronasal organ (VNO) into the brain asserting control of pubertal onset and fertility. Recent data suggest that correct development of the olfactory ensheathing cells (OEC) is imperative for normal GnRH-1 neuronal migration. However, the full ensemble of molecular pathways that regulate OEC development remains to be fully deciphered. Loss-of-function of the transcription factor Gli3 is known to disrupt olfactory development, however, if Gli3 plays a role in GnRH-1 neuronal development is unclear. By analyzing Gli3 extra-toe mutants (Gli3Xt/Xt), we found that Gli3 loss-of-function compromises the onset of achaete-scute family bHLH transcription factor 1 (Ascl-1) positive vomeronasal progenitors and the formation of OEC in the nasal mucosa. Surprisingly, GnRH-1 neurogenesis was intact in Gli3Xt/Xt mice but they displayed significant defects in GnRH-1 neuronal migration. In contrast, Ascl-1null mutants showed reduced neurogenesis for both vomeronasal and GnRH-1ns but less severe defects in OEC development. These observations suggest that Gli3 is critical for OEC development in the nasal mucosa and subsequent GnRH-1 neuronal migration. However, the non-overlapping phenotypes between Ascl-1 and Gli3 mutants indicate that Ascl-1, while crucial for GnRH-1 neurogenesis, is not required for normal OEC development. Since Kallmann syndrome (KS) is characterized by abnormal GnRH migration, we examined whole exome sequencing data from KS subjects. We identified and validated a GLI3 loss-of-function variant in a KS individual. These findings provide new insights into GnRH-1 and OECs development and demonstrate that human GLI3 mutations contribute to KS etiology.Significance statementThe transcription factor Gli3 is necessary for correct development of the olfactory system. However, if Gli3 plays a role in controlling GnRH-1 neuronal development has not been addressed. We found that Gli3 loss-of-function compromises the onset of Ascl1+ vomeronasal progenitors, formation of olfactory ensheathing cells in the nasal mucosa and impairs GnRH-1 neuronal migration to the brain. By analyzing Ascl1 null mutants we dissociated the neurogenic defects observed in Gli3 mutants from lack of olfactory ensheathing cells in the nasal mucosa, moreover, we discovered that Ascl1 is necessary for GnRH-1 ontogeny. Analyzing human whole exome sequencing data, we identified a GLI3 loss-of-function variant in a KS individual. Our data suggest that GLI3 is a candidate gene contributing to KS etiology.

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High-yield production of L-serine through a novel identified exporter combined with synthetic pathway in Corynebacterium glutamicum

10.21203/rs.3.rs-16998/v2 ◽

2020 ◽

Author(s):

Xiaomei Zhang ◽

Yujie Gao ◽

Ziwei Chen ◽

Guoqiang Xu ◽

Xiaojuan Zhang ◽

...

Keyword(s):

Corynebacterium Glutamicum ◽

Fluorescent Protein ◽

Transcriptional Regulator ◽

High Yield ◽

Market Demand ◽

Mobility Shift ◽

Synthetic Pathway ◽

Key Enzyme ◽

Novel Target ◽

Green Fluorescent

Abstract Background: L-Serine has wide and increasing applications in industries with fast-growing market demand. Although strategies for achieving and improving L-serine production in Corynebacterium glutamicum (C. glutamicum) have focused on inhibiting its degradation and enhancing its biosynthetic pathway, L-serine yield has remained relatively low. Exporters play an essential role in the fermentative production of amino acids. To achieve higher L-serine yield, L-serine export from the cell should be improved. In C. glutamicum, ThrE, which can export L-threonine and L-serine, is the only identified L-serine exporter so far.Results: In this study, a novel L-serine exporter NCgl0580 was identified and characterized in C. glutamicum ΔSSAAI (SSAAI), and named as SerE (encoded by serE). Deletion of serE in SSAAI led to a 56.5% decrease in L-serine titer, whereas overexpression of serE compensated for the lack of serE with respect to L-serine titer. A fusion protein with SerE and enhanced green fluorescent protein (EGFP) was constructed to confirm that SerE localized at the plasma membrane. The function of SerE was studied by peptide feeding approaches, and the results showed that SerE is a novel exporter for L-serine and L-threonine in C. glutamicum. Subsequently, the interaction of a known L-serine exporter ThrE and SerE was studied, and the results suggested that SerE is more important than ThrE in L-serine export in SSAAI. In addition, probe plasmid and electrophoretic mobility shift assays (EMSA) revealed NCgl0581 as the transcriptional regulator of SerE. Comparative transcriptomics between SSAAI and the NCgl0581 deletion strain showed that NCgl0581 is a positive regulator of NCgl0580. Finally, by overexpressing the novel exporter SerE, combined with L-serine synthetic pathway key enzyme serAΔ197, serC, and serB, the resulting strain presented an L-serine titer of 43.9 g/L with a yield of 0.44 g/g sucrose, which is the highest L-serine titer and yield reported so far in C. glutamicum. Conclusions: This study provides a novel target for L-serine and L-threonine export engineering as well as a new global transcriptional regulator NCgl0581 in C. glutamicum.

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