\u414\u432\u430 \u41d\u43e\u432\u438 \u411\u435\u437\u441\u435\u440\u443\u43c\u43d\u438 \u418 \u411\u435\u437 \u41f\u440\u43e\u442\u435\u438\u43d\u43d\u438 \u429\u430\u43c\u430, \u41f\u43e\u43b\u443\u447\u435\u43d\u438 \u41e\u442 \u41a\u43b\u435\u442\u44a\u447\u43d\u430 \u41b\u438\u43d\u438\u44f Hep-2: \u41a\u443\u43b\u442\u443\u440\u430\u43b\u43d\u438 \u423\u441\u43b\u43e\u432\u438\u44f \u418 \u41f\u440\u43e\u43b\u438\u444\u435\u440\u430\u442\u438\u432\u43d\u430 \u410\u43a\u442\u438\u432\u43d\u43e\u441\u442

Two new cell strains \u2013 HEp-2-Plovdiv E and HEp-2-Plovdiv F were derived from the HEp-2 cell line. They could be cultivated indefinitely in vitro in DMEM/Ham\u2019s F12 1:1 medium. These are serum-free and protein-free cultures because they grown only in medium that is chemically defined. Serum-free cultures could be distinguished by morphological characteristics, growth dynamics, sensitivity to trypsinization, prolife-ration activity and doubling time. The number of cell population\u2019s doublings of the initial carcinoma cell line and the two serum-free cultures was calculated. The dynamics of cell proliferation was investigated and doubling time for HEp-2 cell line (22.6 h), HEp-2-Plovdiv F strain (25.8 h) and HEp-2-Plovdiv E strain (28.4 h) was calculated. DMSO of 7 % was added in the growth medium for storage in liquid nitrogen. Optimized parameters for cultivation and subcultivation of the cell cultures are suggested. Possible ways of application of the new serum free cell cultures, based on preliminary studies for viral cultivation and detection of serum autoantibodies are discussed.\u41e\u442 \u43a\u43b\u435\u442\u44a\u447\u43d\u430 \u43b\u438\u43d\u438\u44f HEp-2 \u441\u430 \u43f\u43e\u43b\u443\u447\u435\u43d\u438 \u434\u432\u430 \u43d\u43e\u432\u438 \u43a\u43b\u435\u442\u44a\u447\u43d\u438 \u449\u430\u43c\u430 HEp-2-Plovdiv E \u438 HEp-2-Plovdiv F, \u43a\u43e\u438\u442\u43e \u43c\u43e\u433\u430\u442 \u434\u430 \u441\u435 \u43a\u443\u43b\u442\u438\u432\u438\u440\u430\u442 \u43d\u435\u43e\u433\u440\u430\u43d\u438\u447\u435\u43d\u43e \u432\u440\u435-\u435 \u432 \u443\u441\u43b\u43e\u432\u438\u44f \u438\u43d \u432\u438\u442\u440\u43e \u432 \u441\u440\u435\u434\u430 DMEM/Ham\u2019s F-12 (1:1). \u422\u435 \u441\u430 \u431\u435\u437\u441\u435\u440\u443\u43c\u43d\u438 \u438 \u431\u435\u437 \u43f\u440\u43e\u442\u435\u438\u43d\u43d\u438 \u43a\u443\u43b\u442\u443\u440\u438, \u437\u430\u449\u43e\u442\u43e \u441\u435 \u43e\u442\u433\u43b\u435\u436\u434\u430\u442 \u441\u430\u43c\u43e \u432 \u441\u440\u435\u434\u430, \u43a\u43e\u44f\u442\u43e \u435 \u445\u438\u43c\u438-\u447\u435\u441\u43a\u438 \u434\u435\u444\u438\u43d\u438\u440\u430\u43d\u430. \u411\u435\u437\u441\u435\u440\u443\u43c\u43d\u438\u442\u435 \u43a\u443\u43b\u442\u443\u440\u438 \u441\u435 \u440\u430\u437\u43b\u438\u447\u430\u432\u430\u442 \u43f\u43e \u43c\u43e\u440\u444\u43e\u43b\u43e\u433\u438\u447\u43d\u438 \u445\u430\u440\u430\u43a\u442\u435\u440\u438\u441\u442\u438\u43a\u438, \u434\u438\u43d\u430\u43c\u438\u43a\u430 \u43d\u430 \u440\u430\u437\u432\u438\u442\u438\u435 \u43d\u430 \u43a\u443\u43b\u442\u443\u440\u430\u442\u430, \u447\u443\u432\u441\u442\u432\u438\u442\u435\u43b\u43d\u43e\u441\u442 \u43f\u440\u438 \u442\u440\u438\u43f\u441\u438\u43d\u438\u437\u438\u440\u430\u43d\u435, \u43f\u440\u43e\u43b\u438\u444\u435\u440\u430\u442\u438\u432\u43d\u430 \u430\u43a\u442\u438\u432\u43d\u43e\u441\u442 \u438 \u432\u440\u435\u43c\u435 \u43d\u430 \u443\u434\u432\u43e\u44f\u432\u430\u43d\u435. \u418\u437\u447\u438\u441-\u43b\u435\u43d\u438 \u441\u430 \u431\u440\u43e\u44f\u442 \u443\u434\u432\u43e\u44f\u432\u430\u43d\u438\u44f \u43d\u430 \u43a\u43b\u435\u442\u44a\u447\u43d\u430\u442\u430 \u43f\u43e\u43f\u443\u43b\u430\u446\u438\u44f \u437\u430 \u438\u437\u445\u43e\u434\u43d\u430\u442\u430 \u441\u435\u440\u443\u43c\u43d\u430 \u43a\u430\u440\u446\u438\u43d\u43e\u43c\u43d\u430 \u43a\u43b\u435\u442\u44a\u447\u43d\u430 \u43b\u438\u43d\u438\u44f \u438 \u434\u432\u435\u442\u435 \u43d\u43e\u432\u438 \u431\u435\u437\u441\u435\u440\u443\u43c\u43d\u438 \u43a\u443\u43b\u442\u443\u440\u438, \u437\u430\u43b\u43e\u436\u435\u43d\u438 \u432 \u448\u435\u441\u442 \u440\u430\u437\u43b\u438\u447\u43d\u438 \u43d\u430\u447\u430\u43b\u43d\u438 \u43f\u43e\u441\u435\u432\u43d\u438 \u433\u44a\u441\u442\u43e\u442\u438. \u41f\u440\u43e\u441\u43b\u435\u434\u435\u43d\u430 \u435 \u434\u438\u43d\u430\u43c\u438\u43a\u430\u442\u430 \u43d\u430 \u43f\u440\u43e-\u43b\u438\u444\u435\u440\u430\u442\u438\u432\u43d\u430\u442\u430 \u430\u43a\u442\u438\u432\u43d\u43e\u441\u442 \u438 \u435 \u438\u437\u447\u438\u441\u43b\u435\u43d\u43e \u432\u440\u435\u43c\u435\u442\u43e \u43d\u430 \u443\u434\u432\u43e\u44f\u432\u430\u43d\u435 \u43d\u430 \u43a\u43b\u435\u442\u44a\u447-\u43d\u430\u442\u430 \u43a\u443\u43b\u442\u443\u440\u430 HEp-2 (22,6 \u447\u430\u441\u430), \u449\u430\u43c HEp-2-Plovdiv F (25,8 \u447\u430\u441\u430) \u438 \u449\u430\u43c HEp-2-Plovdiv E (28,4 \u447\u430\u441\u430). \u421\u44a\u445\u440\u430\u43d\u44f\u432\u430\u43d\u435\u442\u43e \u432 \u442\u435\u447\u435\u43d \u430\u437\u43e\u442 \u43d\u430 \u431\u435\u437\u441\u435\u440\u443\u43c\u43d\u438\u442\u435 \u43a\u43b\u435\u442\u43a\u438 \u43e\u442 \u434\u432\u430\u442\u430 \u449\u430\u43c\u430 \u441\u435 \u43e\u441\u44a\u449\u435\u441\u442\u432\u44f\u432\u430 \u432 \u440\u430\u441\u442\u435\u436\u43d\u430 \u441\u440\u435\u434\u430, \u434\u43e\u43f\u44a\u43b\u43d\u435\u43d\u430 \u441 7 % DMSO. \u41f\u440\u435\u43f\u43e\u440\u44a\u447\u432\u430\u442 \u441\u435 \u43e\u43f\u442\u438\u43c\u438\u437\u438\u440\u430\u43d\u438 \u43f\u430\u440\u430\u43c\u435\u442\u440\u438 \u43f\u440\u438 \u43f\u440\u43e\u446\u435\u434\u443\u440\u438\u442\u435 \u43d\u430 \u43a\u443\u43b\u442\u438\u432\u438\u440\u430\u43d\u435 \u438 \u441\u443\u431\u43a\u443\u43b\u442\u438\u432\u438\u440\u430\u43d\u435 \u43d\u430 \u43a\u43b\u435\u442\u44a\u447\u43d\u438\u442\u435 \u43a\u443\u43b\u442\u443\u440\u438. \u414\u438\u441\u43a\u443\u442\u438\u440\u430\u442 \u441\u435 \u43d\u44f\u43a\u43e\u438 \u432\u44a\u437\u43c\u43e\u436\u43d\u438 \u43d\u430\u43f\u440\u430\u432\u43b\u435\u43d\u438\u44f \u437\u430 \u43f\u440\u438\u43b\u43e\u436\u435\u43d\u438\u435 \u43d\u430 \u43d\u43e\u432\u438\u442\u435 \u431\u435\u437\u441\u435\u440\u443\u43c\u43d\u438 \u43a\u43b\u435\u442\u43a\u438, \u431\u430\u437\u438\u440\u430\u43d\u438 \u43d\u430 \u43f\u440\u435\u434-\u432\u430\u440\u438\u442\u435\u43b\u43d\u438 \u438\u437\u441\u43b\u435\u434\u432\u430\u43d\u438\u44f \u437\u430 \u43a\u443\u43b\u442\u438\u432\u438\u440\u430\u43d\u435 \u43d\u430 \u432\u438\u440\u443\u441\u438 \u438 \u434\u435\u442\u435\u43a\u446\u438\u44f \u43d\u430 \u441\u435\u440\u443\u43c\u43d\u438 \u430\u432\u442\u43e\u430\u43d\u442\u438\u442\u435\u43b\u430

Два Нови Безсерумни И Без Протеинни Щама, Получени От Клетъчна Линия Hep-2: Културални Условия И Пролиферативна Активност

Two new cell strains – HEp-2-Plovdiv E and HEp-2-Plovdiv F were derived from the HEp-2 cell line. They could be cultivated indefinitely in vitro in DMEM/Ham’s F12 1:1 medium. These are serum-free and protein-free cultures because they grown only in medium that is chemically defined. Serum-free cultures could be distinguished by morphological characteristics, growth dynamics, sensitivity to trypsinization, prolife-ration activity and doubling time. The number of cell population’s doublings of the initial carcinoma cell line and the two serum-free cultures was calculated. The dynamics of cell proliferation was investigated and doubling time for HEp-2 cell line (22.6 h), HEp-2-Plovdiv F strain (25.8 h) and HEp-2-Plovdiv E strain (28.4 h) was calculated. DMSO of 7 % was added in the growth medium for storage in liquid nitrogen. Optimized parameters for cultivation and subcultivation of the cell cultures are suggested. Possible ways of application of the new serum free cell cultures, based on preliminary studies for viral cultivation and detection of serum autoantibodies are discussed.От клетъчна линия HEp-2 са получени два нови клетъчни щама HEp-2-Plovdiv E и HEp-2-Plovdiv F, които могат да се култивират неограничено вре-е в условия ин витро в среда DMEM/Ham’s F-12 (1:1). Те са безсерумни и без протеинни култури, защото се отглеждат само в среда, която е хими-чески дефинирана. Безсерумните култури се различават по морфологични характеристики, динамика на развитие на културата, чувствителност при трипсинизиране, пролиферативна активност и време на удвояване. Изчис-лени са броят удвоявания на клетъчната популация за изходната серумна карциномна клетъчна линия и двете нови безсерумни култури, заложени в шест различни начални посевни гъстоти. Проследена е динамиката на про-лиферативната активност и е изчислено времето на удвояване на клетъч-ната култура HEp-2 (22,6 часа), щам HEp-2-Plovdiv F (25,8 часа) и щам HEp-2-Plovdiv E (28,4 часа). Съхраняването в течен азот на безсерумните клетки от двата щама се осъществява в растежна среда, допълнена с 7 % DMSO. Препоръчват се оптимизирани параметри при процедурите на култивиране и субкултивиране на клетъчните култури. Дискутират се някои възможни направления за приложение на новите безсерумни клетки, базирани на пред-варителни изследвания за култивиране на вируси и детекция на серумни автоантитела

Biological effects of a novel intestinal peptide -- inhibiting enterocytogenin on cultured 3T3 mouse fibroblasts and L5178Y mouse lymphoma cells

The effects of a new intestinal peptide, inhibiting enterocytogenin (IEG) derived from pig intestinal mucosa were studied in vitro on 3T3 mouse fibroblasts and L5178Y mouse lymphoma cell line. IEG caused considerable growth inhibition together with specific morphological changes, necrotic effects as well as formation of monolayers at the highest concentration applied (1000 [mu]g/ml). A biologically active fraction (IEG-BAF) derived by further purification of IEG by gel-filtration, proved to possess most of the described activity. The concentrations of IEG and IEG-BAF inhibiting the growth of L5178Y lymphoma cells by 50% (IC50 values) were calculated to be 759 [mu]g/ml and 192 [mu]g/ml, respectively. IEG-BAF has a molecular mass of 4450 +/- 180 Da and is most probably a peptidylnucleotidate as revealed by spectral analysis.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31582/1/0000511.pd

Experimental Investigations: Micromeritic Procedures In Assessing Antinuclear Antibody Patterns In Immunofluorescent Assay

ЦЕЛЬ: Работа ставит себе целью ввести микрометрическую процедуру (статистический анализ, применя- емый для небольших объектов) в индиректный флюоресцентный анализ в целях получения объективных количественных параметров при анализе локализации антинуклеарных антител в диагностике аутоим- мунных заболеваний. МАТЕРИАЛЫ И МЕТОДЫ: Применены: сыворотки пациентов с системными аутоим- мунными заболяваниями; безсывороточная клеточная линия McCoy-Plovdiv; маркированный флюорохро- мом FITC иммуноглобулин; флюоресцентный микроскоп; цифровизирование и обработка изображений; микрометрический анализ; ANOVA статистическая обработка данных. РЕЗУЛЬТАТЫ: Три вида локализации антиген-антитела (ANA) реакции - гомогенная, периферическая и петнистая - анализированы микроме- трическим методом. Считается, что яркость пикселов флюоресцентных изображений служит объектив- ной характеристикой вида локализации. Максимальный объем и поверхностная плотность связанных с субстратом ANA получены для петнистого типа локализации. ЗАКЛЮЧЕНИЕ: Mикрометрический метод количественной характеристики типа ANA локализации в индиректной иммунофлюоресценции может оказаться ценным дополнительным инструментом для анализа в иммунологических диагностических ла- бораториях с возможностью для включения в пакет программного обепечения (sofware) компьютерной программы для обработки изображений