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Comparison of impact of two decontamination solutions on the viability of the cells in human amnion

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Abstract

Human amniotic membrane (HAM) is used as an allograft in regenerative medicine or as a source of pluripotent cells for stem cell research. Various decontamination protocols and solutions are used to sterilize HAM before its application, but little is known about the toxicity of disinfectants on HAM cells. In this study, we tested two decontamination solutions, commercial (BASE·128) and laboratory decontamination solution (LDS), with an analogous content of antimycotic/antibiotics for their cytotoxic effect on HAM epithelial (EC) and mesenchymal stromal cells (MSC). HAM was processed in a standard way, placed on nitrocellulose scaffold, and decontaminated, following three protocols: (1) 6 h, 37 °C; (2) 24 h, room temperature; (3) 24 h, 4 °C. The viability of EC was assessed via trypan blue staining. The apoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). The mean % (±SD) of dead EC (%DEC) from six fresh placentas was 12.9 ± 18.1. Decontamination increased %DEC compared to culture medium. Decontamination with BASE·128 for 6 h, 37 °C led to the highest EC viability (81.7%). Treatment with LDS at 24 h, 4 °C resulted in the lowest EC viability (55.9%) in the set. MSC were more affected by apoptosis than EC. Although the BASE·128 expresses lower toxicity compared to LDS, we present LDS as an alternative decontamination solution with a satisfactory preservation of cell viability. The basic formula of LDS will be optimised by enrichment with nutrient components, such as glucose or vitamins, to improve cell viability.

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Acknowledgements

The research leading to these results has received funding from the Norwegian Financial Mechanism 2009–2014 and the Ministry of Education, Youth and Sports under Project Contract No. MSMT-28477/2014, Project 7F14156. Institutional support was provided by the PRVOUK-P24/LF1/3 and SVV Project No. 260256/2016 programs of Charles University. Authors thank Brigita Vesela, Dana Sadilkova, Zuzana Rejnysova from Institute of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic, for their excellent technical assistance with the Sudan III staining.

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Authors and Affiliations

  1. Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University and General University Hospital in Prague, Ke Karlovu 2, 128 08, Prague, Czech Republic

    Ingrida Smeringaiova, Peter Trosan, Jan Bednar & Katerina Jirsova

  2. Department of Obstetrics and Gynaecology, Second Faculty of Medicine, Charles University, Prague, Czech Republic

    Miluse Berka Mrstinova & Jan Matecha

  3. Department of Transplantation and Tissue Bank, Motol University Hospital, Prague, Czech Republic

    Jan Burkert & Katerina Jirsova

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  1. Ingrida Smeringaiova
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  2. Peter Trosan
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  3. Miluse Berka Mrstinova
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  4. Jan Matecha
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  5. Jan Burkert
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  6. Jan Bednar
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  7. Katerina Jirsova
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Correspondence to Katerina Jirsova.

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Ingrida Smeringaiova, Peter Trosan, Miluse Berka Mrstinova, Jan Matecha, Jan Burkert, Jan Bednar and Katerina Jirsova declares that they have no conflict of interest.

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Smeringaiova, I., Trosan, P., Mrstinova, M.B. et al. Comparison of impact of two decontamination solutions on the viability of the cells in human amnion. Cell Tissue Bank 18, 413–423 (2017). https://doi.org/10.1007/s10561-017-9636-3

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  • DOI: https://doi.org/10.1007/s10561-017-9636-3

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