ПРЕДОПЕРАЦИОННАЯ ПОДГОТОВКА КЛЕТОЧНЫХ ТРАНСПЛАНТАТОВ ЛИМБА ДЛЯ ЛЕЧЕНИЯ ОПТИЧЕСКИХ НЕЙРОПАТИЙ (ЭКСПЕРИМЕНТАЛЬНОЕ ИССЛЕДОВАНИЕ)

Oftal mokhirurgiya

М. Ð¥. Хубецова¹  Ю А. Комах³  Ð¥. Д. Тонаева⁴  Н И. Ланевская⁵  Л. Н. Скуратовская⁵  И. Н. Сабурина⁵  В. С. Репин⁵  С. А. Борзенок⁵  Д С Островский⁵  Н А. Гаврилова⁶ 

[1] Ð“БОУ ВПО «Московский государственный медико-стоматологический университет им А.И. Евдокимова «Минздрава России;ФГБНУ «Научно-исследовательский институт общей патологии и патофизиологии»;ФГБОУ ДПО «Российская медицинская академия последипломного образования» Минздрава России;ГБОУ ВПО «Московский государственный медико-стоматологический университет им А.И. Евдокимова «Минздрава России;ФГАУ «МНТК «Микрохирургия глаза» им. акад. С.Н. Федорова» Минздрава России;ФГБНУ «Научно-исследовательский институт общей патологии и патофизиологии»

关键词: limbal multipotent mesenchymal stem cells;    3D culture;    cell spheroids;    nerve growth factor (NGF);    brain derived growth factor (BDNF);    cell therapy;    neuroprotection;    optic neuropathy;    3D-культуры;    клеточные сфероиды;    фактор роста нервов (ФРН);    нейроростовой фактор головного мозга (НФГМ);    клеточная терапия;    нейропротекция;    оптическая нейропатия;   

DOI  :  10.25276/0235-4160-2017-2-28-36

学科分类：眼科学

来源: M H T K Mikrokhirurgiya Glaza

【 摘 要 】

Purpose . To study experimentally in vitro secretion of the nerve growth factor (NGF) and the brain-derived neurotrophic factor (BDNF) using intact and induced multi-potent mesenchymal limbal stem cells (MSCs) in three-dimensional culture (3D). Material and methods . MSCs were obtained by culturing of limbal fragments, isolated from the cadaveric human donor eyes, according to the medical technology of the S. Fyodorov Eye Microsurgery Federal State Institution. The phenotype of obtained cell culture was studied by the flow cytometry method.Stimulation of secretion of neurotrophic factors was performed via a two-step technique using non-specific activation factors: EGF, hbFGF, N2 additive, dibutyryl cAMP, NRG1-beta 1, PDGF, 3-isobutyl-1- methylxanthine. The 3D-cell spheroids were generated using agarose plates (3D Petri dishes, Microtissue, USA) for three comparative groups where: Group I – control, Group II – with the induction of spheroids at 1 day of cultivation; Group III – with induction of spheroids at 7 days of cultivation. The concentration of NGF and BDNF in the culture medium was studied using the enzyme linked immunosorbent assay (ELISA). Results . The induction of the 3D spheroids of limbal MSCs, that carried out on the 1st and 7th days of incubation, contributes to a significant increase in the production of NGF and BDNF, but subsequently a pronounced reduction in the secretion of these factors is observed. The conducting of an induction leads to a change in the morphology of spheroids: loss of compactness, the emergence of «fringed» (debris). Such changes indicate a non-viability of the obtained 3D-cell spheroids. Conclusion . The cellular spheroids, created from 2D-culture of intact limbal MSCs by the three-dimensional culture method, are capable in sufficient therapeutic concentrations spontaneously to synthesize NGF and BDNF, have the most optimal design for transplantation in extrabulbar and intraocular tissue niches of the eyeball, are a potential source of prolonged secretion of neural basis function in cell treatment of optic neuropathy. 

【 授权许可】

CC BY