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Immunophenotyping of a population of cultured human umbilical cord cells from Wharton's jelly

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1. Title Title of document Immunophenotyping of a population of cultured human umbilical cord cells from Wharton's jelly
2. Creator Author's name, affiliation, country Vladimir E. Chernov; Military Medical Academy; Russian Federation
2. Creator Author's name, affiliation, country Margarita O. Sokolova; Military Medical Academy; Russian Federation
2. Creator Author's name, affiliation, country Arina A. Kokorina; Military Medical Academy; Russian Federation
2. Creator Author's name, affiliation, country Galina I. Pendinen; N.I. Vavilov All-Russian Institute of Plant Genetic Resources; Russian Federation
3. Subject Discipline(s)
3. Subject Keyword(s) сulturing; cytophotometry; explant; human bud; immunofluorescence; mesenchymal stem cells; mucous connective tissue of the umbilical cord; Wharton's jelly
4. Description Abstract

BACKGROUND: Against the background of many existing methods of defect replacement in post-traumatic injuries the methods of repair of damaged tissues based on methods using products of tissue engineering and cultures of artificially cultured human cells are becoming more and more widespread in medical practice. The literature reports weak immunogenic activity of human umbilical cord tissues, which makes these cells promising components of regenerative medicine products. Due to possible errors in explant selection and cell transformation in the process of cultivation, it is necessary to reliably determine the phenotype of cells obtained as a result of tissue explantation and their further cultivation. Thus, the specificity of obtaining multipotent mesenchymal cells from human umbilical cord Warton's stool tissues requires reliable identification of the type of the obtained cells.

AIM: To obtain reliable features of mesenchymal stem cells in the cell population obtained from human umbilical cord stool tissue.

MATERIAL AND METHODS: Methods of cell cultivation, flow cytofluorimetry, immunocytochemistry for determination of surface and intracellular markers of mesenchymal stem cells were used in the study.

RESULTS: In the course of work on the identification of cells of the population obtained by culturing explants from human umbilical cord Wharton stool, the heterogeneity of the type of cells constituting the cell population was established. Most of them are mesenchymal stem cells carrying fluorescent markers CD45, CD73, CD34, CD29, CD90, CD44, CD105, which agrees with the immunophenotype of mesenchymal stem cells defined by the International Society for Cell Therapy. The ratio of the applied markers allows us to refer the population of cells obtained by direct explantation of Varton's gelatin tissue fragments to mesenchymal stem cells. Visual control confirmed the localisation of labelled antibodies on the surface of cultured cells. And also, it was shown that there were no vascular muscle cells in the obtained culture.

CONCLUSION: As a result of experiments on identification of the cells obtained during explantation of Wharton's jelly tissue fragments and their further cultivation, their belonging to mesenchymal stem cells was established by immunofluorescence cytophotometry.

5. Publisher Organizing agency, location Eco-Vector
6. Contributor Sponsor(s) Government of the Russian Federation (VMA.02.06.2022/026 «Эксплант»)
7. Date (DD-MM-YYYY) 31.07.2024
8. Type Status & genre Peer-reviewed Article
8. Type Type Research Article
9. Format File format
10. Identifier Uniform Resource Identifier https://journals.eco-vector.com/RMMArep/article/view/624871
10. Identifier Digital Object Identifier (DOI) 10.17816/rmmar624871
10. Identifier Digital Object Identifier (DOI) (PDF (Eng)) 10.17816/rmmar624871-150518
11. Source Title; vol., no. (year) Russian Military Medical Academy Reports; Vol 43, No 2 (2024)
12. Language English=en en
13. Relation Supp. Files Fig. 1. MSC culture: А, Б — staining with haematoxylin and eosin; В — immunocytochemical staining for the presence of smooth muscle a-actin in the cytoskeleton of cells obtained from umbilical cord, Г — stained rat myometrial cells (415KB)
Fig. 2. Results of immunofluorescence labelling of surface antigens (SD) of mesenchymal cells of human umbilical cord Warton’s stool tissue. А — mesenchymal cell before labelling; Б, В, Г — images of cells labelled with different antibodies: Б — absence of conjugation with antibody (DAPI contrasting); В, Г — presence of conjugation with labelled antibody (236KB)
Fig. 3. Distribution of surface markers in the obtained population of human umbilical cord MSCs: А — CD34, CD44; Б — CD45, CD73; В — CD29, Г — CD90; Д — CD105 (263KB)
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15. Rights Copyright and permissions Copyright (c) 2024 Eco-Vector
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