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Journals / SDÜ Tıp Fakültesi Dergisi / 2023 / Cilt: 30 Sayı: 4

MORPHOMETRIC AND FLUORESCENCE ANALYSIS OF NERVE GROWTH FACTOR-DIFFERENTIATED PC12 CELLS

SİNİR BÜYÜME FAKTÖRÜ İLE FARKLILAŞTIRILMIŞ PC12 HÜCRELERİNİN MORFOMETRİK VE FLORESANS ANALİZİ

Simge Ünay(LOKMAN HEKIM UNIVERSITY, SCHOOL OF MEDICINE)0000-0002-5582-5574
Ferhat Şirinyıldız(AYDIN ADNAN MENDERES UNIVERSITY, SCHOOL OF MEDICINE)0000-0001-8800-9787
Pages
732–737
Publication Date
2023-12-30
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Results The results showed that the length of neurite extensions and dead cell ratio increased depending on NGF doses and incubation time.

📌 3 Key Takeaways

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Objective & Hypothesis:Objective PC12 is a rat pheochromocytoma cell line.
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Key Findings & Data:However, literature studies showing the effect of NGF on neurite extensions formed in PC12 cells are very limited.
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Conclusion & Impact:Results The results showed that the length of neurite extensions and dead cell ratio increased depending on NGF doses and incubation time.
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:Nörit uzunluğu · PC12 hücre hattı · Sinir büyüme faktörü

Abstract

Objective PC12 is a rat pheochromocytoma cell line. These cells characteristically undergo differentiation when cultured with nerve growth factor (NGF). Depending on the dose of NGF, the length of neurite extensions changes. Thanks to this differentiation property, the cells are used in neuroscience and in modeling pathophysiological diseases such as Alzheimer's, Parkinson's, and Amyotrophic Lateral Sclerosis. However, literature studies showing the effect of NGF on neurite extensions formed in PC12 cells are very limited. This study aimed to investigate the effect of NGF on neurite extensions and cell viability depending on dose and incubation time. Materials and Methods In this study, PC12 cells were incubated with 50 ng/ml and 100 ng/ml NGF for 3, 6 and 7 days. The lengths of neurite outgrowths and dead cell ratios were calculated in incubated cells. Results The results showed that the length of neurite extensions and dead cell ratio increased depending on NGF doses and incubation time. When NGF incubation times were compared, no difference was found between 50 ng/ml NGF 6 days and 100 ng/ml NGF 3 days groups. Conclusion When the dead cell ratios and sizes of neurite extensions in the experimental groups are evaluated, it is thought that 100 ng/ml NGF and 3 days incubation time parameters are ideal for PC12 cell differentiation.

Özet

Amaç PC12 bir sıçan feokromositoma hücre hattıdır. Bu hücreler, sinir büyüme faktörü (NGF) ile kültürlendiğinde karakteristik olarak farklılaşmaya uğramaktadır. NGF dozuna bağlı olarak, nörit uzantılarının uzunlukları değişir. Bu farklılaşma özelliği sayesinde hücreler nörobilimde ve Alzheimer, Parkinson, Amyotrofik Lateral Skleroz gibi patofizyolojik hastalıkların modellenmesinde kullanılmaktadır. Ancak NGF’nin PC12 hücrelerinde oluşan nörit uzantıları üzerine olan etkisini gösteren literatür çalışmaları oldukça kısıtlıdır. Bu çalışmanın amacı NGF’nin doza ve inkübasyon süresine bağlı olarak nörit uzantıları ve hücre canlılığı üzerine olan etkisini araştırmaktır. Gereç ve Yöntem Bu çalışmada PC12 hücreleri 50 ng/ml ve 100 ng/ml NGF ile 3, 6 ve 7 gün inkübe edilmiştir. İnkübe edilen hücrelerde nörit büyümelerinin uzunlukları ve ölü hücre oranları hesaplanmıştır. Bulgular Elde edilen bulgular ile NGF dozlarına ve inkübasyon süresine bağlı olarak nörit uzantılarının uzunluğunun ve ölü hücre oranının arttığı gösterilmiştir. NGF inkübasyon süreleri karşılaştırıldığında 50 ng/ml NGF 6 gün ve 100 ng/ml NGF 3 gün grupları arasında fark olmadığı bulunmuştur. Sonuç Deney gruplarında ölü hücre oranları ve nörit uzantılarının boyutları değerlendirildiğinde 100 ng/ml NGF ve 3 gün inkübasyon süresi parametrelerinin PC12 hücre farklılaşması için ideal olduğu düşünülmektedir.

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Cite This Article

Ünay, S., Şirinyıldız, F. (2023). MORPHOMETRIC AND FLUORESCENCE ANALYSIS OF NERVE GROWTH FACTOR-DIFFERENTIATED PC12 CELLS. SDÜ Tıp Fakültesi Dergisi, Cilt: 30 Sayı: 4, 732–737. https://doi.org/10.17343/sdutfd.1329845

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References (18)

  1. Tang XP, Guo XH, Geng D, Weng LJ. d-Limonene protects PC12 cells against corticosterone- induced neurotoxicity by activating the AMPK pathway. Environ Toxicol Pharmacol. 2019;70:103192. doi:10.1016/j.etap.2019.05.001.
  2. Lee J, Song K, Huh E, Oh MS, Kim YS. Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from Tussilago farfara. Redox Biol. 2018;18:6–15. doi:10.1016/j.redox.2018.05.015.
  3. Yin Z, Han Z, Hu T, et al. Neuron-derived exosomes with high miR-21-5p expression promoted polarization of M1 microglia in culture. Brain Behav Immun. 2020;83:270–282. doi:10.1016/j. bbi.2019.11.004.
  4. Stefanescu R, Stanciu GD, Luca A, et al. Contributions of mass spectrometry to the identification of low molecular weight molecules able to reduce the toxicity of amyloid-β peptide to cell cultures and transgenic mouse models of Alzheimer’s disease. Molecules 2019;24(6):1167.
  5. Lahiani A, Brand-Yavin A, Yavin E, Lazarovici P. Neuroprotective effects of bioactive compounds and MAPK pathway modulation in "Ischemia"- Stressed PC12 pheochromocytoma cells. Brain Sci. 2018;8. doi:10.3390/brainsci8020032.
  6. Wiatrak B, Kubis-Kubiak A, Piwowar A, Barg E. PC12 cell line: cell types, coating of culture vessels, differentiation and other culture conditions. Cells. 2020;9(4):958.
  7. Wang Y, Lai X, Wu D, et al. Umbilical mesenchymal stem cell-derived exosomes facilitate spinal cord functional recovery through the miR-199a-3p/145-5p-mediated NGF/TrkA signaling pathway in rats. Stem Cell Res Ther. 2021;12(1):117. doi:10.1186/s13287-021-02148-5.
  8. Yu C, Cao CY, Shi PD, et al. Highly oxygenated chemical constitutes and rearranged derivatives with neurotrophic activity from Ganoderma cochlear. J Ethnopharmacol. 2022;295:115393. doi:10.1016/j.jep.2022.115393.
  9. Ferreira RS, Dos Santos NAG, Martins NM, Fernandes LS, Dos Santos AC. Caffeic Acid Phenethyl Ester (CAPE) Protects PC12 Cells from Cisplatin-Induced Neurotoxicity by Activating the NGF-Signaling Pathway. Neurotox Res. 2018;34(1):32–46. doi:10.1007/s12640-017-9849-z.
  10. Mirza FJ, Zahid S. The Role of Synapsins in Neurological Disorders. Neurosci Bull. 2018;34(2):349–358. doi:10.1007/ s12264-017-0201-7.
  11. García-Cruz C, Merino-Jiménez C, Aragón J, et al. Overexpression of the dystrophins Dp40 and Dp40L170P modifies neurite outgrowth and the protein expression profile of PC12 cells. Sci Rep. 2022;12(1):1410. doi:10.1038/s41598-022-05271-2.
  12. Molnár J, Frank É, Minorics R, et al. A click approach to novel D-ring-substituted 16α-triazolylestrone derivatives and characterization of their antiproliferative properties. PLoS One 2015;10(2):e0118104. doi:10.1371/journal.pone.0118104.
  13. Ding XW, Li R, Geetha T, Tao YX, Babu JR. Nerve growth factor in metabolic complications and Alzheimer's disease: Physiology and therapeutic potential. Biochim Biophys Acta Mol Basis Dis. 2020;1866(10):165858. doi:10.1016/j.bbadis.2020.165858.
  14. Shilo D, Cohen G, Blumenfeld A, et al. Tuftelin Is Required for NGF-Induced Differentiation of PC12 Cells. J Mol Neurosci. 2019;68(1):135–143. doi:10.1007/s12031-019-01292-1.
  15. Orlowska A, Perera PGT, Al Kobaisi M, et al. The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells. Materials 2017;11:60.
  16. Hu R, Cao Q, Sun Z, et al. A novel method of neural differentiation of PC12 cells by using Opti-MEM as a basic induction medium. Int J Mol Med. 2018;41(1):195–201. doi:10.3892/ ijmm.2017.3195.
  17. Unay S, Bilgin MD. Investigation of effects of quercetin and low-level laser therapy in cisplatin-induced in vitro peripheral neuropathy model. Lasers Med Sci. 2023;38(1):49. doi:10.1007/s10103-023-03718-0.
  18. Sun X, Li W, Gong X, et al. Investigating the Regulation of Neural Differentiation and Injury in PC12 Cells Using Microstructure Topographic Cues. Biosensors 2021;11(10):399. doi:10.3390/ bios11100399.