培养的人脐带粘膜结缔组织（Wharton）细胞群的免疫表型

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论证。在创伤后缺损替代方法层出不穷的背景下，利用组织工程产品和人工培养的人体细胞修复受损组织的方法在医疗实践中越来越广泛。文献报道，人类脐带组织的免疫原性很弱，这使得这些细胞成为再生医学产品中很有前途的成分。由于细胞在培养过程中可能发生自发转化，以及外植体选择过程中的误差，因此有必要对组织外植体及其进一步培养过程中获得的细胞表型进行可靠的鉴定。因此，从人体脐带粘膜结缔组织（也称为Wharton果冻）中获得多能间充质细胞的特异性需要对获得的细胞类型进行可靠的鉴定。

目的。利用免疫酶标记法检测从人类脐带粘膜结缔组织中获取的细胞群中是否存在间充质干细胞。

材料和方法。该研究采用细胞培养、流式细胞荧光测定法和免疫细胞化学法来确定间充质干细胞的表面和细胞内标志物。

结果。在鉴定从人体脐带粘膜结缔组织培养中获得的群体细胞的工作中，确定了构成群体的细胞类型的异质性。它们中的大多数都带有荧光标记 CD45、CD73、CD34、CD29、CD90、CD44、CD105，这与国际细胞治疗学会（International Society for Cell Therapy）定义的间充质干细胞的免疫表型一致。根据应用标记物的比例，我们可以将粘膜结缔组织片段直接外植获得的细胞群归属于间充质干细胞。肉眼观察证实了标记抗体在培养细胞表面的定位。结果还表明，培养物中没有血管肌肉细胞。

结论。通过对粘膜结缔组织碎片外植并进一步培养获得的细胞进行鉴定实验，通过免疫荧光细胞光度法确定其属于间充质干细胞。

作者简介

Vladimir E. Chernov

Military Medical Academy

编辑信件的主要联系方式.
Email: vmeda-nio@mail.ru
ORCID iD: 0000-0002-2440-3782
SPIN 代码: 8315-1161

Cand. Sci. (Biology)

俄罗斯联邦, Saint Petersburg

Margarita O. Sokolova

Military Medical Academy

Email: vmeda-nio@mail.ru
ORCID iD: 0000-0002-3457-4788
SPIN 代码: 3683-6054
俄罗斯联邦, Saint Petersburg

Arina A. Kokorina

Military Medical Academy

Email: arina.alexandrovna.vmeda-nio@mail.ru
ORCID iD: 0000-0002-6783-3088
SPIN 代码: 9371-3658
俄罗斯联邦, Saint Petersburg

Galina I. Pendinen

N.I. Vavilov All-Russian Institute of Plant Genetic Resources

Email: pendinen@mail.ru
ORCID iD: 0000-0003-2814-7074
SPIN 代码: 2120-5925

Cand. Sci. (Biology)

俄罗斯联邦, Saint Petersburg

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2. 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

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3. 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

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4. 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

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