Öz
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.
Anahtar Kelimeler
Proje Numarası
-
Kaynakça
- 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.
Öz
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.
Anahtar Kelimeler
Destekleyen Kurum
-
Proje Numarası
-
Teşekkür
-
Kaynakça
- 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.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Hücre Fizyolojisi |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Proje Numarası | - |
| Yayımlanma Tarihi | 30 Aralık 2023 |
| Gönderilme Tarihi | 19 Temmuz 2023 |
| Kabul Tarihi | 27 Aralık 2023 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 30 Sayı: 4 |
Süleyman Demirel Üniversitesi Tıp Fakültesi Dergisi/Medical Journal of Süleyman Demirel University is licensed under Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International.