مهندسی شیمی ایران

مهندسی شیمی ایران

جداسازی سلول‌های B با استفاده از نانوذرات مغناطیسی و بررسی اثر رنگ‌آمیزی سلول‌ها و مقدار نانوذرات بر جداسازی آن‌ها

نوع مقاله : مقاله پژوهشی

نویسنده
استادیار مهندسی پزشکی، دانشگاه آزاد اسلامی واحد شیراز، شیراز، ایران
چکیده
امروزه از نانوذرات و میکروذرات مغناطیسی در کاربردهای مختلف پزشکی و درمان مانند MRI، ترمیم بافت، رهایش دارو، هایپرترمی و جداسازی سلول‌ها استفاده میشود. یکی از مهم‌ترین سلول‌هایی که باید جداسازی شود، سلول‌های لنفاوی است؛ زیرا نقش بسیار مهمی در درمان سرطان‌های لنفوم و ایمونوتراپی دارد. لذا، در این مطالعه سلول‌های B از PBMCs بااستفادهاز نانوذرت مغناطیسی اکسید آهن پوششدادهشده با پکتین جدا شدهاست. بهمنظور جداسازی هدفمند سلولهای B، آنتیبادی FITC anti-human CD20 بااستفادهاز EDC/NHS به نانوذرات، متصل و با فلوسایتومتری آنالیز شدهاست. در مرحلۀ بعد، سلول‌های B با نانوذرات حاوی آنتیبادی از بقیۀ سلول‌ها جدا و با فلوسایتومتری بازده جداسازی بررسی شدهاست. نتایج فلوسایتومتری
نشان می‌‌‌‌‌دهد که آنتی‌بادی به نانوذرات متصل شده و میزان
MFI آن برابر با 55 است. بازده جداسازی سلول‌ها 75 تا 82 درصد است. هم‌چنین، مشخص شد که با رنگ‌آمیزی سلول‌ها بعد از جداسازی، میزان بازده واکنش تا حدودی نسبتبه حالت بدون رنگ‌آمیزی و آنالیز مستقیم سلول‌ها بعد از شست‌وشو با فلوسایتومتری از 81 درصد به 76 درصد کاهش مییابد. علاوهبر این، نتایج نشان داد که مقدار مناسب نانوذرات در واکنش با سلول‌ها 50 میکرولیتر است و کاهش آن موجب کاهش 30 درصدی در بازده جداسازی می‌شود.
کلیدواژه‌ها

موضوعات


عنوان مقاله English

Isolation of B Cells Using Magnetic Nanoparticles and Investigating the Effect of Cell Staining and the Amount of Nanoparticles on their Isolation

نویسنده English

A. H. Haghighi
Assistant Professor of Biomedical Engineering, Shiraz Branch, Islamic Azad University, Shiraz, Iran
چکیده English

Today, magnetic nanoparticles and microparticles are used in various medical and therapeutic applications such as MRI, tissue repair, drug release, hyperthermia, and cell isolation. One of the most important cells to isolate are lymphoid cells, because they play a very important role in the treatment of lymphoma cancers and immunotherapy. Therefore, in this study, B cells were isolated from PBMCs using iron oxide magnetic nanoparticles coated with pectin. In order to isolate B cells, FITC
anti-human CD20 antibody was attached to nanoparticles using EDC/NHS and analyzed by flow cytometry. In the next step, the target cells are separated from others cells with nanoparticles containing antibodies, and the rate of isolation is checked by flow cytometry.

The results of flow cytometry show that the antibody is attached to the nanoparticles and its MFI is equal to 55. It was also found that by staining the cells after isolation, the reaction efficiency is somewhat reduced from 81% to 76% compared to the state without staining and direct analysis of the cells after washing by flow cytometry. In addition, the results showed that the appropriate amount of nanoparticles in the reaction with cells is 50 microliters and its reduction causes a 30% loss in the separation efficiency.

کلیدواژه‌ها English

Magnetic Nanoparticles
Pectin
B Cells
Flow Cytometry Test
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