Abstract
Giriş: Böbrek hücrelerinin elde edilmesinde kullanılan primer kültür yöntemi, böbrek hastalıklarının hücre düzeyinde değerlendirilmesi ve hücre fonksiyonlarının incelenmesi için olanak sağlamaktadır. Bu çalışmanın amacı böbrek primer kültürü için kullanılabilecek üç farklı izolasyon yönteminin hücre sağkalımı ve protol etkinliği açısından birbirleriyle karşılaştırılmasıdır.
Gereç ve Yöntemler: Nefrektomi ile çıkarılan böbrek dokusu steril şartlarda mekanik, tripsin ve kollagenaz yöntemi ile ayrımlanarak podosit vasatı içerisinde kültüre edildi. Kültüre edilen hücrelerin podosin, vimentin, E-kaderin, TGF-β1, CD133, ZO-1 ve Nefrin dağılımlarının analizi immünositokimyasal yöntem ile yapıldı.
Bulgular: Kollajenaz enzimatik yöntemi sonrasında hücrelerin kültürün 7. gününden itibaren, mekanik yöntemde ise 13. günden itibaren epiteloid karakter aldıkları gözlenirken, tripsin enzimatik yönteminde 13. günde kültürde artan fuziform yapıdaki hücreler gözlendi. Her üç yöntem ile böbrek dokusundan elde edilen podosit hücrelerin immunositokimyasal analizi sonrasında, elde edilen hücrelerde vimentin, E-kaderin, TGF-β, CD133, ZO-1 immunoreaktiviteleri farklılık gösterir iken, podosin ve nefrin immunoreaktivitelerinin kollajenaz enzimatik yöntemi ile elde edilen hücrelerde kuvvetli olduğu saptandı.
Tartışma: Bu çalışmada, farklı proteinlerin immunositokimyasal boyamaları ile kontrol edilen böbrek hücreleri için üç farklı primer kültür teknikleri karşılaştırıldığında, kollajenaz ile muamele edilen böbrek dokusundan daha yüksek verimle hücre saflaştırılabildiği gözlendi. Podosit hücrelerinin kültüre edilmesinde hücre sağkalım verimi ve protokol etkinliği açısından kollajenaz yönteminin en uygun yöntem olduğu gösterildi.
Keywords
References
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Abstract
References
- 1.Chawla, Lakhmir S., and Paul L. Kimmel. "Acute kidney injury and chronic kidney disease: an integrated clinical syndrome." Kidney international 82.5 (2012): 516-524.
- 2. Levey, A,S, Cassandra, B, B, Lesley, A, I, Glomerular filtration rate and albuminuria for detection and staging of acute and chronic kidney disease in adults: a systematic review, Jama, 2015, 313(8), 837-846.
- 3. Ding, W, Keyvan, Y, Lina, A,S, Isolation, characterization, and high throughput extracellular flux analysis of mouse primary renal tubular epithelial cells, Journal of visualized experiments JoVE, 2018, 136.
- 4. Vander, A,J, Sherman J,H, Luciano D,S, The mechanism of body function. In: Vander’s human physiology, McGraw-Hill Education, 2000, pp 476-486.
- 5. Detrisac, C,J, Sens M,A, Garvin A,J, Spicer S,S, Sens D,A, Tissue culture of human kidney epithelial cells of proximal tubule origin, Kidney international ,1984, 25(2), 383-390.
- 6. Uysal O, Sevimli T, Sevimli M, Gunes S, Sariboyaci AE, Cell and Tissue Culture: The Base of Biotechnology. In: Omics Technologies and Bio-Engineering Towards Improving Quality of Life, 1st edn, Academic Press, London, 2018, pp 391-429.
- 7. Hendijani, F, Explant culture: An advantageous method for isolation of mesenchymal stem cells from human tissues, Cell proliferation, 2017,50(2), e12334.
- 8. Valente, M,J, Henrique, R, Costa, V,L, Jerónimo, C, Carvalho, F, Bastos, M,L, Pinho, P,G, Carvalho, M, A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens. PloS one, 2011, 6(5), e19337.
- 9. Hawksworth G,M, Isolation and culture of human renal cortical cells with characteristics of proximal tubules. Methods in Molecular Medicine, 2005, 107, 283–290.
- 10. Vesey, D,A, Qi, W, Chen, X, Pollock, C,A, Johnson, D,W, Isolation and primary culture of human proximal tubule cells. Methods in Molecular Biolology, 2009, 466, 19–24.
- 11. Glynne, P, Primary culture of human proximal renal tubular epithelial cells. Methods in Molecular Medicine, 2000, 36: 197–205.
- 12. Qian, T, Hernday, S,., Bao, X, Olson, W,R, Panzer, S,E, Shusta, E,V, Palecek, S,P, Directed differentiation of human pluripotent stem cells to podocytes under defined conditions, Scientific reports,2019, 9(1), 1-12.
- 13. Satelli, A, Li, S, Vimentin in cancer and its potential as a molecular target for cancer therapy. Cellular and molecular life sciences, 2011, 68(18), 3033-3046.
- 14. Shen, S,S, Krishna, B, Chirala, R, Amato, R,J, Truong, L,D, Kidney-specific cadherin, a specific marker for the distal portion of the nephron and related renal neoplasms. Modern pathology,2005, 18(7), 933-940.
- 15. Sureshbabu, A, Muhsin, S,A, Choi, M,E, TGF-β signaling in the kidney: profibrotic and protective effects. American Journal of Physiology-Renal Physiology, 2016, 310(7), F596-F606.
- 16. Bruno, S., Bussolati, B., Grange, C., Collino, F., Graziano, M. E., Ferrando, U., & Camussi, G. (2006). CD133+ renal progenitor cells contribute to tumor angiogenesis. The American journal of pathology, 169(6), 2223-2235.
- 17. Shankland, S. J., Pippin, J. W., Reiser, J., & Mundel, P. (2007). Podocytes in culture: past, present, and future. Kidney international, 72(1), 26-36.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Biochemistry and Cell Biology (Other) |
| Journal Section | Araştırma Makalesi |
| Authors | |
| Publication Date | September 30, 2021 |
| Published in Issue | Year 2021 Volume: 8 Issue: 3 |
