Genes & CellsGenes & CellsГены и КлеткиGenes and Cells2313-18292500-2562Human Stem Cells Institute12060610.23868/gc120606ArticlesСтатьиResearch ArticleOptimization of teratoma formation assayОптимизация тератомного тестаKizilovaE. AКизиловаЕ. Аpinus@bionet.nsc.ruFederal Research Center Institute of Cytology and Genetics, the Siberian Branch of the Russian Academy of SciencesФГБНУ «Федеральный исследовательский центр Институт цитологии и генетики СО РАН»15062016112VOL 11, NO2 (2016)ТОМ 11, №2 (2016)11912805012023Copyright ©; 2016, Eco-VectorCopyright ©; 2016, Эко-Вектор2016Eco-VectorЭко-Векторhttps://genescells.ru/2313-1829/article/view/120606

Teratoma formation assay is necessary to estimate in vivo pluripotency of stem cells especially stem cell lines of human origin. Nevertheless convenient, valid and universal “standards” to analyze stem cell derived tumors have not been developed yet. New protocol for monitoring teratoma growth, morphological and histological analyzes of tumor samples is proposed in this paper. This protocol is oriented on review of tumors morphology and histology per se. The list-describer includes 17 obligate and 12 facultative diagnostic sell types and 7 diagnostic cell complexes. The protocol takes into account complicity and heterogeneity of teratoma structure and allows detect different morphological features of malignization process inside stem cell derived tumors in situ. The protocol was successfully applied for teratoma formation test which has been performed for 52 stem cell lines of different species origin (mouse, rat, аmerican mink and human). 326 stem cell derived tumors were completely described, reviewed and analyzed.

Сообщение посвящено критическому обзору и анализу тератомного теста, применяемого для оценки плюрипотентности стволовых клеток млекопитающих в условиях in vivo. Особое значение он имеет для анализа стволовых клеток человека. Сегодня клеточные биологи работают над стандартизацией и оптимизацией тератомного теста. Предлагается схема оптимизации, которая учитывает специфику морфологии и гистологии опухолей, происходящих из введённых стволовых клеток. Особое внимание уделено мониторингу роста опухоли, отбору проб, прочтению препаратов и формализации процедуры их описания. Выбраны 17 основных и 12 добавочных клеточных морфотипов, которые надежно распознаются на гистологических препаратах опухоли. Выделены и описаны 7 основных типов клеточных ансамблей, образующихся в опухолях in situ. Предложены критерии для оценки сложности организации опухоли и критерии малигнизации. Схема прошла успешную проверку более чем в 30 долговременных экспериментах. Проведен анализ 326 опухолей, полученных из суспензий 52 линий различных типов стволовых клеток 4 биологических видов (человек, лабораторная мышь, крыса, американская норка). Предложенный алгоритм позволяет полнее и точнее проводить диагностику экспериментальных тератом и, следовательно, увеличивает валидность экспертного заключения о плюрипотентности стволовых клеток в условиях in vivo.

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