Владимир, вы работали с культурами нейрональных клеток и гепатоцитов ?
Гепатоциты и ХеЛя - любимые вирусологами объекты, работал. С нейронами - не работал.
С культурами кое-что уже наработали:
Poster Session
Evaluation of Cancer-derived Exosomes by FT-IR Spectroscopy
Mara Grube1, Karlis Shvirksts1, Arturs Abols2, Aija Line2
1Institute of Microbiology and Biotechnology, University of Latvia, Jelgavas str 1, Riga, LV1004, Latvia
2 Latvian Biomedical Research and Study centre, Ratsupites str 1, LV1067, Riga, Latvia
Exosomes are small nano-sized (30–100 nm) membrane vesicles of endosomal origin released in the extracellular space by a variety of cell types including epithelial cells, mesenchymal stem cells, immune cells and various cancer cells, and are found in all human body fluids. Cancer cells abnormally secrete large quantities of exosomes that transport onco-proteins and immune suppressive molecules thus promoting the growth of tumour and metastasis. Exosomes are composed of proteins, lipids - phospholipids, cholesterol, sphingolipids, ceramides and raft molecules, carbohydrates - mannose, polylactosamine, α-2,6 sialic acid and complex N-glycan’s, which are important to bind proteins in exosome, and nucleic acids such as miRNA, small non-coding RNA and mRNA derived from their cellular origin [1, 2]. In our study FT-IR spectroscopy was used to estimate the purity of extracted exosomes and to evaluate the composition of exosomes produced by colorectal cancer cell lines derived from a primary (SW480) and metastatic (SW620) tumour, which were cultured under hypoxic or normoxic conditions. FTIR spectra were collected using microplate reader HTS-XT (Bruker, Germany). Certainly it was easy to assess the step-by-step purification, to discriminate the fractions of proteins and exosomes and hereafter control the purity of extracted exosomes by FT-IR spectra analyses. In the fraction of purified exosomes the main absorption bands identified proteins (1550-1660 cm-1), lipids (2880-2950 cm-1) and carbohydrates (925-1111cm-1). Yet the content of nucleic acids was too low for precise detection. The composition of exosomes released by SW480 and SW620 cells significantly differed by the content of proteins. The shift in carbohydrate absorption maximum from 1090 cm-1 in normoxic to 1100 cm-1 in hypoxic conditions was observed for both exosome samples. It was shown that interrelations of the protein, lipid and carbohydrate content in exosomes varies depending on their origin and is affected by hypoxia.
References
[1] A.V. Vlassov, S. Magdaleno, R. Setterquist, Conrad Biochim Biophys Acta 1820, 940-948 (2012).
[2] Y. Liang, W.S. Eng, D.R. Colquhoun, R.R. Dinglasan, D.R. Graham, L.K. Mahal, J Biol Chem. 289, 32526-32537 (2014).
Acknowledgment: This study was supported by the LCS joint project ″Cancer-derived exosomes – a source of novel biomarkers and therapeutic targets for gastrointestinal cancers″.
А культура нервных клеток с устойчивой генерацией ПД в конкретном нейроне и потом снятие гликанового портрета с такого нейрона - задача технически очень сложная.
Да, сложная. Их на мультиэлектродных матрицах культивируют и меченными лектинами красят.
Было сообщение, дисс. немца из Тайваня, что степень фукозилирования при шизофрении повышается. Ишемия, или ещё что, нейроны, или что-то другое - не понятно.
Это не значит, что Ваша работа не имеет ценности и не нужна вообще.
Лилия! Я удивляюсь Вашей наивности. Как Вы представляете себе процесс производста заушников? А потом заушники, с нетренированными мозгами, становятся рецензентами.
За время безвремения научная нива покрылась плотной порослью сорняков, теперь они хотят удобрений (т.е.увеличения финансирования). Се ля ви, без селекции элита перестает быть таковой.
