Sel’skokhozyaistvennaya Biologiya [Agricultural Biology], 2018, Vol. 53, ¹ 4, p. 868-875.
(how to cite this article)

doi: 10.15389/agrobiology.2018.4.868eng

UDC 619+616.5]:57.085.23

 

MULTIPOTENT MESENCHYMAL STROMAL CELLS ISOLATED
FROM SUBCUTANEOUS FAT OF MAMMALS FOR THE STUDY
OF Sarcoptes Scabiei/mange in vitro

I.P. Savchenkova

Federal Science Center Kovalenko All-Russian Research Institute of Experimental Veterinary RAS, Federal Agency of Scientific Organizations, 24/1, Ryazanskii pr., Moscow, 109428 Russia, e-mail s-ip@mail.ru (✉ corresponding author)

ORCID:
Savchenkova I.P. orcid.org/0000-0003-3560-5045
The authors declare no conflict of interests

Received October 19, 2017

 

Sarcoptes scabiei/mange is a small, roundish, pale gray mite that lives in the epidermis of mammalian skin and causes scabies. Despite the fact that the mite biology is well studied and its interaction with the host is intensively investigated, the data analysis demonstrates the absence of a cell culture on which the Sarcoptes scabiei/mange could multiply in vitro. This hinders the development of modern effective methods of diagnosis of this disease, the study of the immune response after infection in order to create vaccines and evaluate the effectiveness of the use of drugs for the clinical treatment of mite-borne disease. In this regard, the search for a cellular system that would maintain the viability of Sarcoptes scabiei/mange in vitro is particularly relevant. We describe cellular systems represented by multipotent mesenchymal stromal cells (MMSCs) that can be used for these purposes. MMSCs were isolated from subcutaneous adipose tissue (SAT) of cattle and humans. Biopsies were taken from healthy donors without clinical form of scabies. In MMSC of cattle and human, isolated from SAT scabies mite (Sarcoptes scabiei/mange) was found. The MMSCs isolated from cattle were the most contaminated by mites: of 13 MMSC cultures on the 2nd passage 9 cultures (or 69 %) were contaminated. For human MMSCs, contamination with mites was only found in one culture among 6 cultures obtained (16 %). The mite was found in the form of small dark spherical extracellular components, not characteristic for MMSCs, the number of which increased during culture. A characteristic feature of scabies mite reproduction in MMSC culture isolated from both cattle and human SAT was the formation of nests in the form of clusters. In the culture, we identified presumably larvae and nymphs of the mite. During subculturing within 3 passages, the contaminated cattle and human MMSCs retained the same vitality as not contaminated, but the monolayer formed more slowly, i.e. on days 9-10 vs. day 7 in the control groups. After freezing the samples in which the mite was found, and storage in a Dewar vessel in liquid nitrogen for a week, followed by defrosting, it was found that the mite retained viability in all samples and well tolerated freezing-thawing. After thawing, the mite was detected on the 2nd passage in 7 of 7 analyzed MMSC cultures from cattle SAT and in the only MSCS culture derived from human SAT in which the mite contamination was found (100 %). Thus, it is shown that the MMSCs isolated by us from cattle and humans SAT, may represent a promising cellular system for studying in vitro Scabiei scabieis/mange. Possible contamination of MMSCs from SAT with scabies makes testing of these cultures mandatory in case of their use in cellular technologies.

Keywords: multipotent mesenchymal stromal cells, subcutaneous adipose tissue, cell culture, Sarcoptes scabiei/mange, culturing, freezing.

 

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