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犬乳腺恶性肿瘤原代细胞培养的建立:一项初步研究。

Establishment of primary cell cultures from canine mammary gland malignant tumours: a preliminary study.

作者信息

Petrouškova Patŕicia, Hudáková Nikola, Almášiová Viera, Valenčáková Alexandra, Horňáková L'ubica, Huniadi Mykhailo, Čížková Daša

机构信息

Department of Epizootiology, Parasitology and Protection of One Health, University of Veterinary Medicine and Pharmacy in Košice, 041 81 Košice, Slovak Republic.

Small Animal Clinic, University Veterinary Hospital, University of Veterinary Medicine and Pharmacy in Košice, 041 81 Košice, Slovak Republic.

出版信息

J Vet Res. 2025 Mar 1;69(1):159-168. doi: 10.2478/jvetres-2025-0007. eCollection 2025 Mar.

Abstract

INTRODUCTION

Canine mammary gland cancer (CMGC) is the most common neoplastic condition in bitches and is often fatal. There are limited treatment options for CMGC. Primary cell cultures from mammary tumours are promising preclinical models in which to study personalised treatment approaches. This preliminary study aimed to establish primary cell cultures from two canine mammary gland neoplasms: a common solid adenocarcinoma and a rare carcinosarcoma.

MATERIAL AND METHODS

Tumour masses were collected from a 13-year-old and a 16-year-old German shepherd. Tumour cells were isolated by mechanical disaggregation and enzymatic digestion of masses with 0.05% type IV collagenase. Primary cell cultures were validated by immunocytochemistry for specific markers including mucin 1 (MUC1), cytokeratin 8 and 18 (CK8/18) and Kiel 67 (Ki-67).

RESULTS

Primary cell cultures achieved confluency by day 7 of culture, displaying polygonal cellular morphology. Cultures of both cell types exhibited strong positivity for MUC1 of >99% and high Ki-67 proliferation activity of 43.1% ± 0.5% in the solid adenocarcinoma-derived positive cells and 87.9% ± 2.7% in the carcinosarcoma-derived positive cells. Positivity was observed for CK8/18 of 98.1% ± 0.3% in cells derived from solid adenocarcinoma and 31.6% ± 1.5% in cells derived from carcinosarcoma.

CONCLUSION

With further characterisation, the primary cell cultures established in this study can be expected to show considerable potential as foundational models for cancer research.

摘要

引言

犬乳腺肿瘤(CMGC)是母犬最常见的肿瘤疾病,通常是致命的。CMGC的治疗选择有限。乳腺肿瘤的原代细胞培养是有前景的临床前模型,可用于研究个性化治疗方法。本初步研究旨在从两种犬乳腺肿瘤中建立原代细胞培养:一种常见的实性腺癌和一种罕见的癌肉瘤。

材料与方法

从一只13岁和一只16岁的德国牧羊犬身上采集肿瘤块。通过机械解离和用0.05%IV型胶原酶消化肿瘤块来分离肿瘤细胞。通过免疫细胞化学对包括粘蛋白1(MUC1)、细胞角蛋白8和18(CK8/18)以及Kiel 67(Ki-67)在内的特定标志物对原代细胞培养进行验证。

结果

原代细胞培养在培养第7天达到汇合,呈现多边形细胞形态。两种细胞类型的培养物对MUC1均表现出>99%的强阳性,实性腺癌来源的阳性细胞中Ki-67增殖活性高,为43.1%±0.5%,癌肉瘤来源的阳性细胞中为87.9%±2.7%。实性腺癌来源的细胞中CK8/18阳性率为98.1%±0.3%,癌肉瘤来源的细胞中为31.6%±1.5%。

结论

随着进一步的表征,本研究中建立的原代细胞培养有望作为癌症研究的基础模型显示出巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e94/11936083/bf3118a89989/j_jvetres-2025-0007_fig_001.jpg

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