Влияние трансплантации мезенхимных стволовых клеток на течение экспериментального инсульта у крыс
Головной мозг, в отличие от других органов, обладает особенностями структурно-функциональной организации, что в первую очередь объясняется существованием внутримозговых барьеров. В экспериментах на животных проведены трансплантации клеточного материала различных типов (фетальных нейронов головного мозга, фетальных нейронов, культивированных in vitro в нейросферах, а также сингенных или… Читать ещё >
Содержание
- Актуальность проблемы
- Цели и задачи исследования
- Основные положения, выносимые на защиту
- Научная новизна работы
- Теоретическое и практическое значение работы
- Апробация работы
- Глава 1. Аналитический обзор литературы
- 1. 1. Ранние изменения при ишемии головного мозга
- 1. 2. Воспаление и глиальная реакция
- 1. 2. 1. Воспаление
- 1. 2. 2. Глиальная реакция
- 1. 3. Реорганизация ткани после ишемического инсульта. Ограниченная репарация в головном мозге
- 1. 4. Зона ишемической полутени
- 1. 5. Современные методы лечения инсульта
- 1. 5. 1. Улучшение перфузии
- 1. 5. 2. Нейропротекция
- 1. 5. 3. Клеточная терапия инсульта 34 1.5.4 Мезенхимные стволовые клетки как агент клеточной терапии '
- 2. 1. Эксперименты in vitro 49 2.1.1 Выделение и культивирование мезенхимных стволовых клеток из стромы костного мозга крыс
- 2. 1. 2. Фенотипирование мезенхимных стволовых клеток крыс
- 2. 1. 3. Дифференцировка in vitro в ортодоксальных направлениях
- 2. 1. 4. Дифференцировка in vitro в нейрональном направлении
- 2. 1. 5. Окрашивание мезенхимных стволовых клеток крыс флуорохромом РКН
- 2. 2. Эксперименты in vivo
- 2. 2. 1. Введение
- 2. 2. 2. Моделирование экспериментального ишемического инсульта головного мозга
- 2. 2. 3. Введение
- 2. 2. 4. Фиксация тканей головного мозга
- 2. 2. 5. Выявление флуоресцентно меченых МСК в тканях головного мозга
- 2. 2. 6. Морфологический анализ тканей головного мозга
- 2. 2. 7. Иммуногистохимическое окрашивание срезов головного мозга
- 2. 2. 8. Морфометрическая оценка объема дефекта головного мозга и количества сосудов в поврежденном и неповрежденном полушарии
- 2. 2. 9. Поведенческое тестирование животных в водном лабиринте Морриса
- 3. 1. Характеристики МСК крысы in vitro
- 3. 1. 1. Фенотипирование МСК
- 3. 1. 2. Дифференцировка МСК в ортодоксальных направлениях in vitro
- 3. 1. 3. Дифференцировка MCK в нейрональном направлении in vitro
- 3. 1. 4. Окрашивание клеток флуорохромом РКН
- 3. 2. Характеристика головного мозга после локального введения МСК в мозг интактных крыс
- 3. 3. Морфологическая характеристика головного мозга крысы после экспериментального ишемического инсульта
- 3. 3. 1. Распределение меченых флуорохромом РКН 26 клеток в ткани мозга
- 3. 3. 2. Характеристика структуры головного мозга ложнооперированных животных
- 3. 3. 3. Характеристика структуры головного мозга после окклюзии средней мозговой артерии (группа контроля)
- 3. 3. 4. Характеристика структуры головного мозга после окклюзии средней мозговой артерии группа клеточной терапии)
- 3. 3. 5. Морфометрическое исследование головного мозга
- 3. 3. 6. Поведенческое тестирование животных
GFP — зеленый флуоресцирующий белок HIF-1 — hypoxia-inducible factor-1 ICAM-1 — intercellular adhesion molecule TL — интерлейкин IL-lra-IL-1 receptor antagonist iNOS — индуцибельная NO-синтаза LFA-1 — leukocyte function-associated antigen MCP-1 — monocyte chemoattractant protein Meis2 и Pbx Tuj 1 иНи
МНС 1 — молекула главного комплекса гистосовместимости первого класса
MRF-1 — microglial response factor
NeuN — маркерный белок нейронов NeuN
NG2 — протеогликан NG
NGF — nerve growth factor
NMDA и АМРА — рецепторы и ингибиторы пресинаптического высвобождения глутамата N0 — оксид азота
PDGF — ростовой фактор тромбоцитов РКН 26 — липофильный краситель РКН rt-PA — рекомбинантный тканевой активатор плазминогена
STAT — signal transducers and activators of transcription
TGF-P — трансформирующий фактор роста (
TNF-a — фактор некроза опухолей
VCAM-1 — vascular cell adhesion molecule
VEGF — фактор роста эндотелия сосудов
VLA-4 — very late antigen
БТШ-70 — белок теплового шока
ВКМ — внеклеточный матрикс
ГСК — гематопоэтические стволовые клетки
ГФКБ — глиальный фибриллярный кислый белок
ГЭБ — гематоэнцефалический барьер
ДАГ — диацилглицерол
И-З-ф — инозитол-три-фосфат
КОЕ — колоний-образующая единица
МСК — мезенхимные стволовые клетки
НСК — нейрональные стволовые клетки
ОСМА — окклюзия средней мозговой артерии
ОСМА — окклюзия средней мозговой артерии
СВЗ — субвентрикулярная зона
СК — стволовые клетки
ФСБ — фосфатно-солевой буфер
ФСК — фетальные стволовые клетки
ЦНС — центральная нервная система
ЭСК — эмбриональные стволовые клетки
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