Detection of the residual concentration of sodium dodecyl sulfate in the decellularized whole rabbit kidney extracellular matrix.

Autor: Ghorbani F; Tracheal Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Ekhtiari M; Pediatric Urology and Regenerative Medicine Research Center, Pediatric Center of Excellence, Department of Pediatric Urology, Children's Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharibs Street, Keshavarz Boulevard, 1419733151, Tehran, Iran., Moeini Chaghervand B; Lung Transplantation Research Center, National Research Institute of Tuberculosis and Lung Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Moradi L; Pediatric Urology and Regenerative Medicine Research Center, Pediatric Center of Excellence, Department of Pediatric Urology, Children's Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharibs Street, Keshavarz Boulevard, 1419733151, Tehran, Iran., Mohammadi B; Pediatric Urology and Regenerative Medicine Research Center, Pediatric Center of Excellence, Department of Pediatric Urology, Children's Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharibs Street, Keshavarz Boulevard, 1419733151, Tehran, Iran., Kajbafzadeh AM; Pediatric Urology and Regenerative Medicine Research Center, Pediatric Center of Excellence, Department of Pediatric Urology, Children's Medical Center, Tehran University of Medical Sciences, No. 62, Dr. Gharibs Street, Keshavarz Boulevard, 1419733151, Tehran, Iran. kajbaf105@gmail.com.
Jazyk: angličtina
Zdroj: Cell and tissue banking [Cell Tissue Bank] 2022 Mar; Vol. 23 (1), pp. 119-128. Date of Electronic Publication: 2021 Apr 28.
DOI: 10.1007/s10561-021-09921-z
Abstrakt: To optimize rabbit kidney decellularization protocol, using sodium dodecyl sulfate (SDS) as a commonly used detergent, a methylene blue based assay was employed for detecting the minimum nontoxic SDS level for future cell seeding. The rabbit kidney tissues were decellularized with the perfusion-based method and underwent several investigations to determine the efficacy of decellularization in preserving the extracellular matrix (ECM) and cell removal. SDS detection was performed by incubating with methylene blue and subsequent extraction with chloroform. MTT (3-(4, 5-dimethylthiazol-2-yr)-2,5-diphenyltetrazolium bromide) assay and SDS release were also evaluated during the entire process. After the first washing cycle, SDS concentration was 0.036, in 500 mL of the washing liquid, which slowly decreased and reached to 0.009 % after at the end of seventh washing cycle. In the 9th cycle, SDS was gradually decreased and reached to 0.003 %. SDS was significantly released after one week of incubation which ceased after ten washing cycles. The results of MTT assay demonstrated that different cells exhibited various sensitivity levels when exposed to serial concentrations of SDS. Human embryonic kidney cells (HEK293) with 0.003 % threshold for cellular toxicity and 87.4 % cell viability were more resistant compared with mesenchymal stem cells with 0.001 % threshold and 85.4 % cell viability. Colorimetric assay with methylene blue is a straightforward and non-invasive method to detect residual SDS present in tissue and can also prevent ECM destruction after several washings for detergent removal from decellularized tissues.
(© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)
Databáze: MEDLINE
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