Изучение особенностей пролиферации эмбрилнальных клеток японских ускоренно стареющих мышей (SAM)
Диссертация
Клетки, которые вышли из митотического цикла на неопределенное время, сохраняя жизнеспособность и пролиферативный потенциал, называют покоящимися клетками (Епифанова и др., 1983). Такие клетки, находящиеся вне митотического цикла («вышедшие из цикла»), содержат, как правило, пресинтетическое количество ДНК. В отличие от фаз цикла их состояние обозначается как GO-фаза, или состояние… Читать ещё >
Содержание
- Список сокращений
- Обзор литературы
- 1. Клеточный цикл 9 1.1 .Состояние пролиферативного покоя
- 1. 2. Факторы роста
- 1. 3. Активация тирозинкиназных рецепторов факторами роста
- 2. Проблема концевой недорепликации
- 2. 1. Пролиферация клеток in vitro. Ограничение пролиферативного 17 потенциала клеток многоклеточных организмов
- 2. 2. «Теломерная теория старения, или теория маргинотомии» А. Оловникова
- 2. 3. Экспериментальные подтверждения теории A.M. Оловникова
- 3. Теломеры
- 3. 1. Теломерный повтор
- 3. 2. Функции теломер
- 4. Теломераза
- 4. 1. РНК — компонент теломеразы
- 4. 2. Теломеразные белки
- 4. 3. Работа теломеразы
- 4. 4. Механизм образования 3' - одноцепочечного фрагмента
- 4. 5. Экзонуклеазная активность теломераз
- 4. 6. Альтернативные механизмы поддержания длины теломер
- 4. 7. Ингибиторы обратных транскриптаз нуклеотидной природы
- 5. Пролиферация клеток различных видов в культуре
- 5. 1. Морфологические стадии старения фибробластов в культуре
- 5. 2. Фибробласты человека в культуре
- 5. 3. Старение, или Ml
- 5. 4. Состояние «кризиса», или М
- 5. 5. Клетки мыши в культуре
- 6. Ускоренно стареющие мыши линий SAMP и SAMR
- 7. Связь между старением клеток в культуре и естественным старением 46 организма
- 7. 1. Корреляция пролиферативного потенциала клеток с видовой 46 продолжительностью жизни
- 7. 2. Зависимость пролиферативного потенциала фибробластов от возраста 46 донора
- 7. 3. Низкий пролиферативный потенциал фибробластов при наследственных 48 болезнях преждевременного старения — прогериях
- 7. 4. Данные об ограниченности потенциала клеточных делений in vivo
- Материалы и методы
- 1. Животные линий SAMP-1, SAMR-1, СВА
- 2. Получение животных с датированной беременностью
- 3. Получение эмбриональных фибробластов мыши
- 4. Условия культивирования клеток
- 5. Замораживание клеточных культур
- 6. Размораживание клеточных культур
- 7. Культивирование клеток в присутствии перекиси водорода
- 8. Выявление активности эндогенной (5-галактозидазы (рН=6,0)
- 9. Пролиферативный покой и стимуляция ростовыми факторами
- 10. Радиоавтография
- 11. Исследование фосфорилирования тирозина
- 12. Определение теломеразной активности (ТАК)
- 13. Ингибиторы обратных транскриптаз
- 14. Изучение телом ер
- 15. Гибридизация in situ
- 16. Приготовление хромосомных препаратов
- Результаты
- 1. Рост эмбриональных фибробластов мышей в культуре
- 2. Количественные параметры роста и старения культур эмбриональных 63 фибробластов мышей линий SAMP-1, SAMR-1 и СВА
- 3. Пролиферативный потенциал ЭФМ линий SAMP-1, SAMR-1 и СВА
- 4. Время переживания клеток в неделящемся состоянии
- 5. Выявление активности эндогенной Р-галактозидазы (рН=6,0)
- 6. Пролиферативный ответ эмбриональных фибробластов мышей на стимуляцию 68 факторами роста
- 7. Уровень фосфорилирования белков по тирозиновым остаткам
- 8. Спонтанная трансформация
- 9. Теломеразная активность (ТАК)
- 10. Подавление функции теломеразы с помощью ингибиторов обратных 74 транскриптаз
- 11. Изучение телом ер и кариотипа эмбриональных фибробластов мышей линий 76 SAMP-1 и SAMR
- Обсуждение
- 1. Морфологически старение клеток SAM неотличимо от старения нормальных 81 мышиных фибробластов
- 2. Повышенная активность эндогенной (3-галактозидазы (рН=6,0) отражает 81 ускоренное старение эмбриональных фибробластов мышей SAM
- 3. Снижение пролиферативного ответа на стимуляцию факторами роста в 81 процессе старения эмбриональных фибробластов мышей в культуре
- 4. Вызывается ли спонтанная трансформация эмбриональных фибробластов 83 мышей в культуре повышением (появлением) ТАК?
- 5. Существует два пути спонтанной трансформации
- 6. Связь длины телом ер у мышей и пролиферативного потенциала их клеток в 85 культуре
- 7. Объяснение того, как относительно длинные теломеры мыши могут 86 индуцировать пролиферативное старение
- 8. Тройственная корреляция между продолжительностью жизни мышей, 88 пролиферативным потенциалом их эмбриональных фибробластов и временем переживания состарившихся фибробластов в неделящемся состоянии
- 9. Повышенная нестабильность ДНК в клетках SAM может объяснить все 91 наблюдаемые в наших экспериментах явления
- Выводы
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