Effects of different culture media on optimization of primary neuronal cell culture for in vitro models assay

Document Type: Reproduction - Physiology

Authors

1 Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Surgery and Radiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

Abstract

Background: In vitro model studies are becoming increasingly popular for experimental research designs. They include isolation and expansion of cells of a particular tissue, such as the nervous tissue which contributes to understanding the underlying mechanisms in many pathologies. It enables  the scrutinization of intracellular signaling pathways responsible for cell death. OBJECTIVES: In the literature, there are different methods for the isolation and culture of rat embryonic cortical neurons. However, this study developed a feasible, rapid and easily performable method. METHODS: Isolation of neurons was performed without using enzymatic digestion. Primary cortical cultures neurite outgrowth and neuron numbers per field of common mediums were compared for neuronal cells isolation and expansion. In this study, three different culture mediums were considered: Medium I: Neurobasal medium, B-27 and L-glutamine; Medium II: DMEM, FBS and L-glutamine; and Medium III: DMEM/F-12, FBS and L-glutamine. RESULTS: High survival rate and number of neurons was obtained with the current method. The best neuronal growth was achieved by Medium I, while Medium II and III had moderate effect on the neurite outgrowth. CONCLUSIONS: Enzyme-free treatment was introduced and Medium I was used as an alternative method for optimal neuron isolation and expansion. The neuronal cultures are similar to nervous tissue in physiological aspects. Hence, Medium I is more similar to the in vivo condition compared to Mediums II and III.

Keywords


Article Title [Persian]

تأثیر محیط کشت های مختلف بر روی بهینه سازی کشت اولیه سلول های عصبی برای ارزیابی مدل های برون تنی

Authors [Persian]

  • محمد حسین گرانمایه 1
  • علی باغبان زاده 1
  • عباس برین 2
  • جمیله سالار آملی 1
  • محمد مهدی دهقان 3
1 گروه علوم پایه، دانشکده دامپزشکی دانشگاه تهران، تهران، ایران
2 گروه میکروبیولوژی، دانشکده دامپزشکی دانشگاه تهران، تهران، ایران
3 گروه جراحی و رادیولوژی، دانشکده دامپزشکی دانشگاه تهران، تهران، ایران
Abstract [Persian]

زمینه مطالعه:  مطالعه مدلهای برونتنی برای طراحی تحقیقات تجربی رو به افزایش است. از جمله این نوع مطالعات جداسازی و کشت سلولها از بافتهای مختلف مانند بافتهای عصبی میباشد، که در شناخت مکانیسمهای اساسی آسیبها میتواند بسیار کمک کننده باشد و ما را قادر میسازد تا مسیرهای سیگنالینگ درون سلولی مسئول مرگ سلولی را به دقت مورد بررسی قرار دهیم. هدف: اگر چه تا کنون، روشهای مختلفی برای کشت نورونهای قشری جنین رت توصیف شده است. با این حال، مطالعه موجود روش عملی و سریعی را معرفی میکند که میتواند در این نوع مطالعات مورد استفاده قرار گیرد. روش کار: در این مطالعه، از سه محیط کشت مختلف، به ترتیبی که توضیح داده میشود برای جداسازی و کشت نورونها در کشت اولیه قشری بدون استفاده از هضم آنزیمی استفاده گردید: محیطهای مورد استفاده عبارت بودند از: محیط I: محیط نوروبازال، B-27 و ال-گلوتامین؛ محیط II: DMEM، FBS و ال-گلوتامین؛ و محیط IIIFBS،ز12-DMEM/F، و ال-گلوتامین. نتایج: میزان زنده مانی نورونی در این مطالعه بسیار قابل توجه بود و بهترین رشد نورونی در محیط I مشاهده گردید، در حالیکه محیطهای II و III اثر متوسطی بر روی رشد نوریت داشتند. نتیجهگیری نهایی: نتایج این مطالعه نشان میدهد محیط I بیشترین تشابه با شرایط برون تنی را در مقایسه با محیطهای II و III ایجاد نموده و سلولهای نورونی بیشتر قادر به حفظ خصوصیات فیزیولوژیک خود در این محیط میباشند.

Keywords [Persian]

  • نورونهای قشری
  • جداسازی
  • کشت اولیه سلول
  • رت
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