Process Development in Isolation and Culture of HumanMonocytes from Cord Blood; Simple and Cos t-EffectiveMethod
- سال انتشار: 1401
- محل انتشار: بیست و سومین کنگره بین المللی هیبریدی پزشکی تولید مثل و هجدهمین کنگره هیبریدی فناوری سلولهای بنیادی رویان
- کد COI اختصاصی: RROYAN23_302
- زبان مقاله: انگلیسی
- تعداد مشاهده: 67
نویسندگان
Department of Applied Cell Sciences, Faculty of Basic Sciencesand Advanced Medical Technologies, Royan Ins titute, ACECR, Tehran,Iran . Department of Regenerative Medicine, Cell Science ResearchCenter, Royan Ins titute for S tem Cell Biology and Technolog
Department of Research and Development, Royan S tem CellTechnology Company, Cord Blood Bank, Tehran, Iran
Department of Regenerative Medicine, Cell Science ResearchCenter, Royan Ins titute for S tem Cell Biology and Technology,ACECR, Tehran, Iran Department of Research and Development, Royan S tem CellTechnology Company, Cord Blood Bank, Tehran, Iran
Department of Regenerative Medicine, Cell Science ResearchCenter, Royan Ins titute for S tem Cell Biology and Technology,ACECR, Tehran, Iran
چکیده
Objective: Macrophages are multifunctional immune cells thatare widely used for immunological research. To s tudy the functionof macrophages in vitro, umbilical cord blood is a convenientand non-invasive source that provides sufficient numbersof monocytes and macrophages. Monocyte isolation proceduresvary widely in terms of purity, yield, cos t, and technical difficulty.We aimed to find an optimized protocol for monocyte isolationfrom cord blood and compare the effect of three differentvariables in monocyte differentiation and culture techniques: ۱.Cell culture media; ۲. Serum sources and ۳. Cell culture systemson cellular phenotype, adhesion, and viability.Materials and Methods: We used double density gradient centrifugationtechniques (Ficoll® and hyperosmotic Percoll®) formonocyte isolation and flow cytometry analysis of light scatteringand/or expression of pan surface markers, such as CD۳,CD۱۴, and CD۱۹ to determine cell viability and purity. Afterpurification, cells were cultured in adhesion preventing tissueculture plates in either RPMI-۱۶۴۰ or DMEM-F۱۲ or IMDMculture medium supplemented with either ۰.۵-۱۰% fetal bovineserum (FBS), human serum (HS), or human platelet lysate(hPL).Results: We introduce a novel, reproducible, and inexpensiveisolation method that yields monocytes with high purity(from۷۰ to ۹۸%). Also, the results showed that monocyte adhesionto the plate is significantly increased in the presence of alow concentration of serum, and IMDM containing ۵% hPL canbe the bes t option for macrophage cultivation and differentiation.Conclusion: This optimized method is a simple and cos t-effectivescale-up method to provide uniform and pure monocytesfor s tudies of the innate defense sys tem.کلیدواژه ها
Cell Viability, Innate Immune Sys tem, Monocyte,Umbilical Cord Bloodاطلاعات بیشتر در مورد COI
COI مخفف عبارت CIVILICA Object Identifier به معنی شناسه سیویلیکا برای اسناد است. COI کدی است که مطابق محل انتشار، به مقالات کنفرانسها و ژورنالهای داخل کشور به هنگام نمایه سازی بر روی پایگاه استنادی سیویلیکا اختصاص می یابد.
کد COI به مفهوم کد ملی اسناد نمایه شده در سیویلیکا است و کدی یکتا و ثابت است و به همین دلیل همواره قابلیت استناد و پیگیری دارد.