scholarly journals Ultrastructural changes in the spinal cord of rats with experimental allergic encephalomyelitis under the influence of human umbilical cord-derived multipotent mesenchymal stromal cells cryopreserved according to different protocols

2021 ◽  
Vol 9 (1) ◽  
Author(s):  
V. Tsymbaliuk ◽  
◽  
V Vaslovych ◽  
L. Pichkur ◽  
L. Liubich ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Spinal Cord ◽  
Stromal Cells ◽  
Fetal Bovine Serum ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Ultrastructural Changes ◽  
Human Umbilical Cord ◽  
Allergic Encephalomyelitis ◽  
Fetal Bovine ◽  
Experimental Allergic

The transplantation of multipotent mesenchymal stromal cells (MMSCs) is considered to be a possible therapy of multiple sclerosis. For the clinical application of human umbilical cord-derived MMSCs (UC-MMSCs) it is necessary to develop a method of their cryopreservation taking into account the type of cryoprotective media and to investigate the possibility of using these cells for therapeutic purposes in vivo. The purpose of the study was to investigate the effect of UC-MMSCs, cryopreserved in solutions of different composition, on the processes of demyelination and remyelination of the spinal cord of rats with experimental allergic encephalomyelitis (EAE) as a model of multiple sclerosis. Materials and methods. The EAE was modeled by subcutaneous administration of homogenized spinal cord of adult rats with complete Freund's adjuvant. On the 18th day rats with moderate relapsing-remitting form of EAE were suboccipitally injected 1•106 UC-MMSCs, cryopreserved in cryoprotective media containing dimethyl sulfoxide (DMSO), fetal bovine serum (FBS), ethylene glycol, trehalose and sucrose at different composition. On the 35th and 60th days, the studies of ultrastructural changes of the lumbar spinal cord (L3-L5) were performed, assessing the degree of demyelination of nerve fibers by the ratio of myelin sheath (MS) thickness to the diameter of the axis cylinder (AC) of axons. Results. In rats with moderate EAE from the 35th to the 60th day after the modelling of the disorder, destructive changes and signs of demyelination in the spinal cord increased; the MS/AC index corresponded to the average degree of axon demyelination. Suboccipitally administered cryopreserved UC-MMSCs to EAE rats, depending on the used cryopreservation solution, slowed or stopped the demyelination, decreased the MS/AC index to a low degree of axonal demyelination. Reducing the concentration of DMSO in the cryopreservation medium from 10 % to 4 % and adding 6 % trehalose provided a better effectiveness of UC-MMSCs in decreasing the degree of demyelination in EAE. At the same time, the standard solution (10 % DMSO, 90 % FBS) provided these effect, but to a lesser extent. The use of a multicomponent cryopreservation medium containing 15 % ethylene glycol, 3 % DMSO, 10 % sucrose, 12 % trehalose and 60 % FBS did not achieve the goal of maintaining the effects of UC-MMSCs to reduce the degree of demyelination in EAE. Conclusions. To maintain the therapeutic properties of UC-MMSCs, it is advisable to add a reduced concentration of DMSO (4 %) and 6 % trehalose to the cryopreservation medium, supplemented with 90 % fetal bovine serum.

scholarly journals Efficiency of Experimental Allergic Encephalomyelitis Modeling as an Experimental Model of Multiple Sclerosis

2021 ◽  
Vol 6 (6) ◽  
pp. 57-65
Author(s):  
O. O. Nefodov ◽  
◽  
Yu. P. Miasoied ◽  
M. V. Solomenko ◽  
O. V. Velykorodna-Tanasiichuk ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Spinal Cord ◽  
Body Weight ◽  
Experimental Allergic Encephalomyelitis ◽  
Rating Scale ◽  
The Body ◽  
Free Access ◽  
Pathological State ◽  
Allergic Encephalomyelitis ◽  
Experimental Allergic

The purpose of the study was the most adequate modeling of multiple sclerosis in the form of autoimmune allergic encephalomyelitis for further use in the study of experimental equivalents of neurodegenerative conditions. Materials and methods. The article highlights the results of the formation of experimental allergic encephalomyelitis, for the induction of which homologous brain homogenates were used, which in terms of encephalitogenicity ranks first among other drugs (homologous, heterogeneous brain and spinal cord homogenates). An encephalitogenic mixture was injected into the connective tissue of the base of the animal's tail at the rate of 0.1 ml per 100 g of body weight. Results and discussion. Experimental allergic encephalomyelitis reproducible by intradermal administration of a homogenate of the brain, spinal cord and peripheral nerves with a Freund's stimulator belongs to a true autoimmune disease of the nervous system and is an experimentally reproducible model of multiple sclerosis. According to the experimental data obtained, the state of the animals showed that the rats of the experimental group lost body weight, had a number of pathological neurological symptoms, which manifested on average from 10-12 days. The increase in symptoms continued for an average of 7 days and was not very stable. Experimental animals had paresis, manifestations of ataxia, walking disorders and urinary and fecal incontinence, lethargy, which corresponded to 1-2 degrees of the rating scale in 13 animals. In 7 rats, paresis of the hind limbs and loss of tail tone were expressed (grade 3). Most of the rats recovered spontaneously within 6-8 days after the onset of clinical manifestations (18-20 days after immunization). According to the requirements of the experiment, during the paralytic stage, the rats had free access to food and water. In addition, our experimental studies on the formation of experimental allergic encephalomyelitis were confirmed morphologically using electron microscopy. Conclusion. The use of this technique for the formation of experimental allergic encephalomyelitis made it possible to obtain a simulated pathological state of multiple sclerosis in the form of experimental allergic encephalomyelitis and can be used in further studies to identify the corresponding patterns, the degree and nature of changes in the immune and nervous systems of the body during the reproduction of an experimental pathological state. The data obtained can serve as a basis for further studies of drugs and their combinations in order to improve and rationalize the pharmacotherapy of multiple sclerosis

scholarly journals Transfer of experimental allergic encephalomyelitis in Lewis rats using supernates of incubated sensitized lymph node cells.

1977 ◽  
Vol 145 (5) ◽  
pp. 1405-1410 ◽  
Author(s):  
C C Whitacre ◽  
P Y Paterson
Keyword(s):  
Spinal Cord ◽  
Lymph Node ◽  
Guinea Pig ◽  
Experimental Allergic Encephalomyelitis ◽  
Nervous Tissue ◽  
Allergic Encephalomyelitis ◽  
Lewis Rats ◽  
Transfer Activity ◽  
Lymph Node Cells ◽  
Experimental Allergic

Supernates derived from incubated lymph node cells of Lewis rats sensitized to guinea pig spinal cord-Freund's adjuvant transfer experimental allergic encephalomyelitis (EAE) to syngeneic recipients. EAE supernatant transfer activity (EAE-STA) is not demonstrable in supernates derived from LNC of control donors not sensitized to nervous tissue. After addition of brain antigen to active supernates, EAE-STA is not longer demonstrable.

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