MSCs ARE LOCATED IN THE PERINEURIUM OF THE RECIPIENT RAT AFTER ALLOTRANSPLANTATION INTO DAMAGED NERVE

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Abstract


The gene and cell therapy, stimulating the regeneration of damaged nerves is currently under development. In these experiments mesenchymal stem cells (MSCs) are often used. The purpose of this study is to describe the localization and morphological features of mesenchymal stem cells derived from bone marrow after their allografting into the damaged rat nerve. MSCs of the bone marrow of Wistar-Kyoto rats were obtained from Transtechnology LLC (Head G.Polyntsev, Ph.D.). MSCs were cultured, identified and labeled by 5-bromo-2’-deoxyuridine (BrdU) in vitro. The sciatic nerve of adult Wistar-Kyoto rats (n = 12) was damaged (ligature, 40 sec), and the suspension of BrdU+ MSCs (5 · 104 cells in 5 μl per animal) was immediately transplanted into the damaged sciatic nerve. In a previous study, we have showed that some transplanted cells are located in the epineurium of the recipient’s nerve. The perineurium of the recipient rats was studied in the present work. Perineurial cells have polygonal form, thin and flat cell nucleus and form several layers, the basement membranes being placed between them. Perineurium is characterized by the presence of occluding junctions that can be identified using anti-claudine antibodies. The use of antibodies to claudine allowed us to identify perineurium. Some BrdU+ MSCs were found to survive 5-7 d following surgery and according to their localization and morphology became perineurium cells. Such extracellular matrix proteins as laminin, fibronectin, collagen are present in the perineurium. Apparently, the presence of these proteins creates a favorable biological environment for MSCs survival and for their differentiation towards perineurium cells. The results of the study confirm the mesenchymal origin of perineurium cells.

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The gene and cell therapy, stimulating the regeneration of damaged nerves is currently under development. In these experiments mesenchymal stem cells (MSCs) are often used. The purpose of this study is to describe the localization and morphological features of mesenchymal stem cells derived from bone marrow after their allografting into the damaged rat nerve. MSCs of the bone marrow of Wistar-Kyoto rats were obtained from Transtechnology LLC (Head G.Polyntsev, Ph.D.). MSCs were cultured, identified and labeled by 5-bromo-2’-deoxyuridine (BrdU) in vitro. The sciatic nerve of adult Wistar-Kyoto rats (n = 12) was damaged (ligature, 40 sec), and the suspension of BrdU+ MSCs (5 · 104 cells in 5 μl per animal) was immediately transplanted into the damaged sciatic nerve. In a previous study, we have showed that some transplanted cells are located in the epineurium of the recipient’s nerve. The perineurium of the recipient rats was studied in the present work. Perineurial cells have polygonal form, thin and flat cell nucleus and form several layers, the basement membranes being placed between them. Perineurium is characterized by the presence of occluding junctions that can be identified using anti-claudine antibodies. The use of antibodies to claudine allowed us to identify perineurium. Some BrdU+ MSCs were found to survive 5-7 d following surgery and according to their localization and morphology became perineurium cells. Such extracellular matrix proteins as laminin, fibronectin, collagen are present in the perineurium. Apparently, the presence of these proteins creates a favorable biological environment for MSCs survival and for their differentiation towards perineurium cells. The results of the study confirm the mesenchymal origin of perineurium cells. Keywords: МСК; периневрий; регенерация нерва.

About the authors

E S Petrova

Institute of Experimental Medicine, Saint Petersburg


E N Isaeva

Institute of Experimental Medicine, Saint Petersburg


E A Kolos

Institute of Experimental Medicine, Saint Petersburg


D E Korzhevskii

Institute of Experimental Medicine, Saint Petersburg


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Copyright (c) 2019 Petrova E.S., Isaeva E.N., Kolos E.A., Korzhevskii D.E.

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