Получение инсулин - продуцирующих клеток из мультипотентных стромальных клеток человека тема диссертации и автореферата по ВАК РФ 03.02.07, кандидат биологических наук Федюнина, Ирина Александровна

• Федюнина, Ирина Александровна
• кандидат биологических наук
• 2011

Оглавление диссертации кандидат биологических наук Федюнина, Ирина Александровна

СПИСОК СОКРАЩЕНИЙ

ГЛАВА 1. ВВЕДЕНИЕ

1.1. Цель и задачи исследования.

1.2. Научная новизна и практическая значимость.

1.3. Положения, выносимые на защиту.Ю

1.4. Апробация работы.

1.5. Публикации.

1.6. Структура и объем диссертации.

ГЛАВА 2. СОВРЕМЕННОЕ СОСТОЯНИЕ ПРОБЛЕМЫ. АНАЛИЗ ЛИТЕРАТУРНЫХ ДАННЫХ

2.1. Развитие поджелудочной железы.

2.2. Генетические механизмы развития поджелудочной железы.

2.3. формирование энтодермы.

2.4. Ключевые гены панкреатической детерминации и дифференцировки.

2.4.1. Ген PDXI.

2.4.2. ГЕНNKX2.2.■„.

2.4.3. ГЕНЫ NKX6.1 HNKX6.2.

2.4.4. PTFlА.

2.5. Гены эндокринной дифференцировки.

2.5.1. Neurod.

2.5.2. Neurogenin 3.

2.5.3. Notch сигнальный путь.

2.5.4. HESl.

2.5.5. ISLl.

2.5.6. РАХ4 ИРЛХб.

2.5.7. MAFA.

2.6. Проблема лечения сахарного диабета.

2.7. сигнальный путь инсулина

2.8. Диабет зрелого типа у молодых (MODY).

2.9. Современные подходы к лечению инсулинзависимого сахарного диабета.

2.9.1. Терапия, основанная на восстановлении регенерации в-клепок островков Лангерганса.

2.9.2. Терапия, основанная на трансплантации островков Лангерганса.

2.9.3. Терапия-шрного диабетапомощьюволовых клеток.

ГЛАВА 3. МАТЕРИАЛЫ И МЕТОДЫ

3.1. Получение культуры клеток.

3.2. Выделение CD146+ популяции клеток.

3.4. Дифференцирование выделенных МСК.

3.5. Получение аденовирусной конструкции.

3.6. Трансфекцпя клеток и индукция панкреатической дифференцировки.

3.7. Анализ экспрессии панкреатических маркерных генов.

3.7.1. Анализ изменения транскрипции генов.

3.7.2. иммуноцитохимическое окрашивание клеток.

3.7.3. Анализ транскрипции инсулина во времени.

3.7.4. ELISA (тест на толерантность к глюкозе).

3.8. Статистическая обработка данных.

ГЛАВА 4. РЕЗУЛЬТАТЫ И ОБСУЖДЕНИЕ

4.1. Трансфекция рАо5-Рбх1 стандартных МСК из жировой ткани и пупочного канатика человека.

4.2. трансфекция ра05-рбх1 МСК периваскулярного фенотипа из жировой ткани и пупочного канатика человека.

4.3. Выбор условий дифференцировки трансфицированных популяций МСК СВ146+СБ31- человека рА1)5-Рох1.

4.4. Анализ экспрессии нестина в исследованных популяциях МСК.

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