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

Olfactory ensheathing cells (OECs) are a promising potential cell therapy to aid regeneration. However, there are significant challenges in isolating and characterizing them. In the current study, we have explored methods to enhance the recovery of cells expressing OEC marker p75NTR from rat mucosa. With the addition of a 24-hour differential adhesion step, the expression of p75NTR was significantly increased to 73 ± 5% and 46 ± 18% on PDL and laminin matrices respectively. Additionally, the introduction of neurotrophic factor NT-3 and the decrease in serum concentration to 2% FBS resulted in enrichment of OECs, with p75NTR at nearly 100% (100 ± 0% and 98 ± 2% on PDL and laminin respectively), and candidate fibroblast marker Thy1.1 decreased to zero. Culturing OECs at physiologically relevant oxygen tension (2–8%) had a negative impact on p75NTR expression and overall cell survival. Regarding cell potency, co-culture of OECs with NG108-15 neurons resulted in more neuronal growth and potential migration at atmospheric oxygen. Moreover, OECs behaved similarly to a Schwann cell line positive control. In conclusion, this work identified key bioprocessing fundamentals that will underpin future development of OEC-based cell therapies for potential use in spinal cord injury repair. However, there is still much work to do to create optimized isolation methods

An overview of tissue engineering approaches for management of spinal cord injuries

Severe spinal cord injury (SCI) leads to devastating neurological deficits and disabilities, which necessitates spending a great deal of health budget for psychological and healthcare problems of these patients and their relatives. This justifies the cost of research into the new modalities for treatment of spinal cord injuries, even in developing countries. Apart from surgical management and nerve grafting, several other approaches have been adopted for management of this condition including pharmacologic and gene therapy, cell therapy, and use of different cell-free or cell-seeded bioscaffolds. In current paper, the recent developments for therapeutic delivery of stem and non-stem cells to the site of injury, and application of cell-free and cell-seeded natural and synthetic scaffolds have been reviewed

Estrogen receptor immunoreactivity in schwann-like brain macroglia

Olfactory ensheathing cells, tanycytes, pituicytes, pineal glia, retinal Muller cells, and Bergmann glia of normal male rats express concomitantly estrogen receptor, low-affinity neurotrophin receptor, antigen O4, and GFAP, markers characteristic of nonmyelinating Schwann cells. These cells were able to survive and proliferate when cultured from adult tissue, promoted neurite outgrowth, and could guide and ensheath growing neurites. We called this distinct group of growth-promoting central nervous system (CNS) macroglia aldynoglia (Greek: to make grow). Its proliferative and growth-promoting properties seem to be retained during the whole lifetime of the organism in those CNS loci where normal function depends on continuous axon renewal. Aldynoglia plasticity seems totally or partially lost with age where and when it is no longer critical, as in the case of adult cortical and spinal cord radial gila. The concomitant expression of estrogen receptor and low-affinity neurotrophin receptor may promote Schwann-like plasticity of glial cells

Spinal implants of olfactory ensheathing cells promote axon regeneration and bladder activity after bilateral lumbosacral dorsal rhizotomy in the adult rat

Purpose: We performed spinal implantation of olfactory ensheathing cells to demonstrate dorsal root afferent regeneration as well as bladder activity restoration after lumbosacral L6 to S2 rhizotomy. Materials and Methods: Spinal segments receiving bladder innervation, usually L6, S1 and S2, were identified by bipolar stimulation of the ventral roots. Bilateral section of the identified dorsal roots L6 to S2 was performed in 18 male Wistar rats. Immediately after rhizotomy olfactory ensheathing cells or vehicle was unilaterally injected in the vicinity of the sacral parasympathetic nucleus in 9 rats each using a glass micropipette and air pulse system. The severed roots were reattached to the cord with fibrin glue and the animals recovered under antibiotic prophylaxis. Results: Anatomical regeneration of bladder wall primary afferents was demonstrated by the presence of labeled wheat germ agglutinin-horseradish peroxidase fibers in the dorsal horn and sacral parasympathetic nucleus in 8 of 9 cases of olfactory ensheathing cell implantation but not in the 9 controls injected with vehicle. One week after surgery all rats had an atonic bladder on cystometrography. At 6 weeks 8 of the 9 olfactory ensheathing cell implanted rats had recovered bladder activity. No recovery was observed in controls, in which vehicle was injected instead of olfactory ensheathing cells. Conclusions: Regenerated primary afferent fibers from the bladder project to the sacral parasympathetic nucleus, where they presumably form synapses mediating the recovery of bladder activity. Thus, olfactory ensheathing cell implants in the adult rat promote sensory axon regeneration, target reinnervation and bladder activity restoration

Ensheathing cell-conditioned medium directs the differentiation of human umbilical cord blood cells into aldynoglial phenotype cells

CONACYT [11-2933 53444]; [194021

Noncommutativity, Cosmology and ?

Despite their similarities to bone marrow precursor cells (PC), human umbilical cord blood (HUCB) PCs are more immature and, thus, they exhibit greater plasticity. This plasticity is evident by their ability to proliferate and spontaneously differentiate into almost any cell type, depending on their environment. Moreover, HUCB-PCs yield an accessible cell population that can be grown in culture and differentiated into glial, neuronal and other cell phenotypes. HUCB-PCs offer many potential therapeutic benefits, particularly in the area of neural replacement. We sought to induce the differentiation of HUCB-PCs into glial cells, known as aldynoglia. These cells can promote neuronal regeneration after lesion and they can be transplanted into areas affected by several pathologies, which represents an important therapeutic strategy to treat central nervous system damage. To induce differentiation to the aldynoglia phenotype, HUCB-PCs were exposed to different culture media. Mononuclear cells from HUCB were isolated and purified by identification of CD34 and CD133 antigens, and after 12 days in culture, differentiation of CD34+ HUCB-PCs to an aldynoglia phenotypic, but not that of CD133+ cells, was induced in ensheathing cell (EC)-conditioned medium. Thus, we demonstrate that the differentiation of HUCB-PCs into aldynoglia cells in EC-conditioned medium can provide a new source of aldynoglial cells for use in transplants to treat injuries or neurodegenerative diseases. " 2012 Japan Human Cell Society and Springer.",,,,,,"10.1007/s13577-012-0044-5",,,"http://hdl.handle.net/20.500.12104/41239","http://www.scopus.com/inward/record.url?eid=2-s2.0-84862118784&partnerID=40&md5=13dc5afb2c358e59da984c736df6caa

Ensheathing cell-conditioned medium directs the differentiation of human umbilical cord blood cells into aldynoglial phenotype cells.

Record Owner: From MEDLINE, a database of the U.S. National Library of Medicine.; Status: MEDLINE; Publishing Model: Journal available in: Print-Electronic Citation processed from: Internet; NLM Journal Code: 8912329, ay1; Registry Number/Name of Substance: 0 (AC133 antigen). 0 (Antigens, CD). 0 (Antigens, CD34). 0 (Culture Media, Conditioned). 0 (Glycoproteins). 0 (Peptides).; Entry Date: 2013030

Subtractive hybridization identifies genes differentially expressed by olfactory ensheathing cells and neural stem cells

CONACYT [J33999N]; AERM; NMTR [170295, 170325

Microarray analysis of striatal embryonic stem cells induced to differentiate by ensheathing cell conditioned media

The mammalian central nervous system contains well-defined regions of plasticity in which cells of the aldynoglia phenotype promote neuronal growth and regeneration. Only now are the factors that regulate the production of new cells from multipotential neural precursors (MNP) starting to be identified. We are interested in understanding how differentiation towards the aldynoglia phenotype is controlled, and to study these events we have induced the differentiation of embryonic MNP towards this phenotype in vitro. Accordingly, we have used microarrays to analyze gene expression in three different cell populations: olfactory bulb ensheathing cells (EC), a prototypic aldynoglia cell type; undifferentiated MNP; and MNP differentiated in vitro for 24 hr in EC-conditioned media. The expression profiles identified support the idea that the EC are more closely related to Schwann cells and astrocytes than to oligodendrocytes. Following MNP differentiation, more strongly expressed genes define a neuroglial cell phenotype. RT-PCR confirms that S100a6, Mtmr2, and Col5a were highly expressed by EC, whereas Pou3f3 were more strongly expressed in MNP than in EC, and SafB1 and Mash1 expression were induced in MNP by EC-conditioned media. The profile of gene expression after differentiation suggests that Wnt signaling may be inactivated during this process, while activation of the BMP pathway may be elicited through the BMPr1A. These results provide us with a starting point to study the genes involved in the induction of aldynoglia differentiation from MNP. � 2008 Wiley-Liss, Inc

Modifications in the seizures susceptibility by excitotoxic neuronal damage and its possible relationship with the pharmacoresistance

The neuronal damage and seizures are two processes closely related not only as cause and effect in reciprocal way but also through the cellular mechanisms and signaling pathways that they share. Therefore, increments in extracellular levels of the glutamate excitatory neurotransmitter, the over-activation of its receptors and the excessive neuronal excitation, have been described as events associated to both processes. In general, if neurons are not able to recover from its excessive excitation, then they die by excitotoxicity. Our group has showed that the excitotoxicity induced by monosodium glutamate in early developmental stages is able to produce significant modifications in glutamatergic and GABAergic neurotransmission systems. Moreover, preliminary results indicate that those modifications are able to increase the seizure susceptibility in the adulthood, particularly when the convulsive drug 4-aminopyridine and the GABA antagonists are employed to induce the seizures, but not when NMDA agonists are used. Through this chapter the topics mentioned above and the hypothesis about the excitotoxic neonatal damage is able to induce a kind of pharmacoresistance to NMDA analogs will be discussed with in detail. © 2013 Springer Science+Business Media, LLC. All rights are reserved