Обоснование применения мезенхимальных стволовых клеток, полученных in vitro у детей с гематологическими и онкологическими заболеваниями

Выводы.

1. Полученные при помощи усовершенствованной методики культивирования in vitro МСК КМ показали возможность приживления при трансплантациях у детей с гематологическими и онкологическими заболеваниями.

2. МСК, полученные в результате усовершенствованной методики культивирования in vitro, сохраняют полноценную функциональную активность, способность к дифференцировке в адипоциты и остеоциты.

3. Для стандартной оценки МСК в трансплантационном материале предложена следующая иммунофенотипическая панель: CD105, CD166, CD44, CD73, CD13, CD29, CD90.

4. При длительном культивировании in vitro МСК КМ человека 3−4 пассажа обладают высокой пролиферативной активностью, которая снижается при дальнейшем культивировании и становится минимальной к 10−12 пассажу.

5. После 3 пассажа в клеточной популяции отсутствовали клетки с гемопоэтическим и эндотелиальном фенотипом (CD45, CD34, CD 133, CD3, CD 19, CD25, CD38, CD45, CD106, CD31). Наблюдалась высокая экспрессия маркеров МСК KM (CD90, CD 105, CD 166, CD44, CD73, промежуточная CD 13 и СТ)29.).При увеличении срока культивирования МСК до 10−12 пассажа снижается количество клеток, экспрессирующих CD90, CD 105 и CD166. Наблюдается тенденция к снижению CD29, CD13, а также исчезают примеси гемопоэтических клеток (CD45+), эндотелиальных клеток (CD31+) и моноцитов (CD 14+).

6. В результате длительного культивирования у МСК КМ на ранних и поздних пассажах хромосомный набор не менялся (46,XY или 46, XX) и отсутствовали анеуплоидии.

7. Полученные нами данные оценки трансфузионной безопасности у детей позволяют предположить, об отсутствии ранних трансфузионных осложнений при применении приготовленных in vitro МСК у детей.

8. Используемый метод ПЦР определения химеризма МСК КМ продемонстрировал достаточную информативность и может применяться для оценки приживления МСК у детей с гематологическими и онкологическими заболеваниями.

Практические рекомендации.

1. Метод может быть рекомендован для экспансии ex vivo МСК КМ в клинических целях.

2. Для культивирования МСК in vitro рекомендованы образцы костного мозга, время у которых от момента забора костного мозга донора до начала выделения и культивирования прошло не более 6 часов.

3. Для оценки иммунофенотипа МСК в трансплантационном материале рекомендуется использовать панель, состоящую из характерных для МСК маркеров (CD90, CD105, CD166, CD44, CD73, CD13, CD29), так и маркеров, специфичных для гемопоэтических клеток (CD3, CD14, CD 19, CD25, CD29, CD31, CD34, CD38, CD45, CD 106, HLA-DR).

4. Для клинических целей рекомендовано применять МСК 3−4 пассажа.

5. В случае необходимости использования в терапевтических целях МСК более поздних сроков культивирования необходим строгий контроль поверхностного фенотипа и генетической стабильности клеточных трансплантатов.

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