Isolation and \(\textit{in vitro}\) cultivation of human urothelial cells from urine of patients

Authors

  • Phan Thi Kieu Trang Institute of Genome Research, VAST, Vietnam
  • Nguyen Thanh Duong Graduate University of Science and Technology, VAST, Vietnam
  • Nguyen Huy Hoang Institute of Genome Research, VAST, Vietnam

DOI:

https://doi.org/10.15625/2615-9023/15915

Keywords:

Bladder cancer, cell culture, mechanical properties, urothelial cells

Abstract

In clinical practice the number of urothelial cells collected by biopsy are limited and the procedure requires general anaesthesia. Therefore, in order to acquire enough urothelial cells for in vitro engineering of the urothelium, in this research we aim to isolate urothelial cells from human urine by an alternative, effective, low-cost and safe technique rather than using the indicated method. Sixty human urine samples had been collected from patients and volunteers, cells then were precipitated by centrifugation and cultured. Following the isolation process, these cells were characterized by the immunocytochemical method using some specific antibodies. There are 2 types of cells were successfully isolated from with different shape and morphology, one of them grew randomly while the others formed smooth-edge contours and cobblestone-like cell morphology. These cells were characterized by immunostaining with a specific marker, both of these cells were positive for urothelial marker cytokeratin 7. All these results were taken into consideration, the isolated cells were urothelial cells observed in the urine-derived cell population. These results will be used for in vitro studies in toxicological and clinical research, and it will be the premised research to determine the cell mechanical properties and then develop a promising method for early diagnosis of bladder cancer.

Downloads

Download data is not yet available.

References

Abidine Y., Laurent V. M., Michel R., Duperray A., Verdier C., 2015. Local mechanical properties of bladder cancer cells measured by AFM as a signature of metastatic potential. Eur. Phys. J. Plus, 130: 202. https://doi.org/10.1140/epjp/ i2015-15202-6

Alibert C., Goud B., Manneville J. -B., 2017. Are cancer cells really softer than normal cells? Biol. Cell, 109: 167–189. https://doi.org/10.1111/boc.201600078

Detrisac C. J., Mayfield R. K., Colwell J. A., Garvin A. J., Sens D. A., 1983. In vitro culture of cells exfoliated in the urine by patients with diabetes mellitus. J. Clin. Invest., 71: 170–173.

Dörrenhaus A., Müller J. I., Golka K., Jedrusik P., Schulze H., Föllmann W., 2000. Cultures of exfoliated epithelial cells from different locations of the human urinary tract and the renal tubular system. Arch. Toxicol., 74: 618–626. https://doi.org/10.1007/s002040000173

Felix, J.S., Littlefield, J.W., 1980. Human newborn urine as a source of epithelial cells. Birth Defects Orig. Artic. Ser. 16: 231–237.

Felix J. S., Littlefield J. W., 1979. Urinary Tract Epithelial Cells Cultured from Human Urine, in: Nichols W. W., Murphy D. G., Toji L. H., Jacobs L. J., Miller R. C. (Eds.), International Review of Cytology, Differentiated Cells in Aging Research. Academic Press, pp. 11–23. https://doi.org/10.1016/S0074-7696(08)60 609-9

Grégoire M., Fradet Y., Meyer F., Têtu B., Bois R., Bédard G., Charrois R., Naud A., 1997. Diagnostic accuracy of urinary cytology, and deoxyribonucleic acid flow cytometry and cytology on bladder washings during followup for bladder tumors. J. Urol., 157: 1660–1664.

Herz F., 1980. Culture of urinary cells. Birth Defects Orig. Artic. Ser., 16: 85–93.

Herz F., Gazivoda P., Papenhausen P. R., Katsuyama J., Koss L. G., 1985. Normal human urothelial cells in culture. Subculture procedure, flow cytometric and chromosomal analyses. Lab. Investig. J. Tech. Methods Pathol., 53: 571–574.

Herz F., Schermer A., Koss L. G., 1979. Short-term culture of epithelial cells from urine of adults. Proc. Soc. Exp. Biol. Med. Soc. Exp. Biol. Med. N. Y. N., 161: 153–157. https://doi.org/10.3181/00379727-161-40509

Inoue C. N., Sunagawa N., Morimoto T., Ohnuma S., Katsushima F., Nishio T., Kondo Y., Iinuma K., 2003. Reconstruction of tubular structures in three-dimensional collagen gel culture using proximal tubular epithelial cells voided in human urine. In Vitro Cell. Dev. Biol. Anim., 39: 364–367.

International Agency for Research on Cancer, For Vietnam, October 2020.

Linder D., 1976. Culture of cells from the urine and bladder washings of adults. Somatic Cell Genet., 2: 281–283. https://doi.org/10.1007/BF01538966

Liu H., Tan Q., Geddie W. R., Jewett M. A. S., Phillips N., Ke D., Simmons C. A., Sun Y., 2014. Biophysical characterization of bladder cancer cells with different metastatic potential. Cell Biochem. Biophys., 68: 241–246. https://doi.org/ 10.1007/s12013-013-9702-9

Lokeshwar V. B., Habuchi T., Grossman H. B., Murphy W. M., Hautmann S. H., Hemstreet G. P., Bono A. V., Getzenberg R. H., Goebell P., Schmitz-Dräger B. J., Schalken J. A., Fradet Y., Marberger M., Messing E., Droller M. J., 2005. Bladder tumor markers beyond cytology: International Consensus Panel on bladder tumor markers. Urologyi, 66: 35–63. https://doi.org/10.1016/j.urology.2005.08.064

Lotan Y., Roehrborn C. G., 2003. Sensitivity and specificity of commonly available bladder tumor markers versus cytology: results of a comprehensive literature review and meta-analyses. Urology, 61: 109–118; discussion 118. https://doi.org/10.1016/s0090-4295(02)02 136-2

Nguyen Ky, Nguyen Buu Trieu, 1993. Results of surgical treatment for bladder cancer for 10 years at Viet Duc hospital. Journal of Surgery, 3: 7−15.

Pavelka M., & Roth J., 2010. Umbrella Cell - Surface Specialisations. Functional Ultrastructure: 250–251. https://doi.org/ 10.1007/978-3-211-99390-3_129

Rahmoune H., Thompson P. W., Ward J. M., Smith C. D., Hong G., Brown J., 2005. Glucose transporters in human renal proximal tubular cells isolated from the urine of patients with non-insulin-dependent diabetes. Diabetes, 54: 3427–3434. https://doi.org/10.2337/diabetes. 54.12.3427

Ramos J. R., Pabijan J., Garcia R., Lekka M., 2014. The softening of human bladder cancer cells happens at an early stage of the malignancy process. Beilstein J. Nanotechnol., 5: 447–457. https://doi.org/ 10.3762/bjnano.5.52

Resnick M. J., Chang S. S., 2013. Optimizing outcomes for octogenarians with invasive bladder cancer: One size does not fit all. Urol. Oncol., 31: 1–4. https://doi.org/ 10.1016/j.urolonc.2011.12.024

Sens M. A., Hintz D. S., Rutledge D. S., Jenkins M. Q., Sens D. A., 1984. Tissue culture of epithelial cells from urine. II. Application to patients with cystic fibrosis. Pediatr Pathol, 2: 165–170.

Sutherland G. R., Bain D., 1972. Culture of cells from the urine of newborn children. Nature, 239: 231.

Wallace D. M. A., Bryan R. T., Dunn J. A., Begum G., Bathers S., West Midlands Urological Research Group, 2002. Delay and survival in bladder cancer. BJU Int., 89: 868–878. https://doi.org/10.1046/ j.1464-410x.2002.02776.x

Zhou T., Benda C., Duzinger S., Huang Y., Li X., Li Y., Guo X., Cao G., Chen S., Hao L., Chan Y. -C., Ng K. -M., Ho J. C., Wieser M., Wu J., Redl H., Tse H. -F., Grillari J., Grillari-Voglauer R., Pei D., Esteban M. A., 2011. Generation of induced pluripotent stem cells from urine. J. Am. Soc. Nephrol. JASN, 22: 1221–1228. https://doi.org/10.1681/ASN.2011 010106

Downloads

Published

20-09-2021

How to Cite

Trang, P. T. K., Duong, N. T., & Hoang, N. H. (2021). Isolation and \(\textit{in vitro}\) cultivation of human urothelial cells from urine of patients. Academia Journal of Biology, 43(3), 19–26. https://doi.org/10.15625/2615-9023/15915

Issue

Section

Articles

Most read articles by the same author(s)

<< < 1 2