Interdisciplinary Research, Central Research Facility, Dr. D. Y. Patil Medical College, Hospital, and Research Center, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, India.
Interdisciplinary, School of Health Sciences, Savitribai Phule, Pune University, India.
Asian Pac J Cancer Prev. 2023 Jan 1;24(1):257-266. doi: 10.31557/APJCP.2023.24.1.257.
Due to many uses of cell culture in cell biology, biotechnology, and medical research, this technique has evolved into a widely used and accepted methodology. The isolation of primary cells from primary cancer tissue is a crucial step in cell culture technology since it offers a trustworthy source for studying the biology, morphology, and molecular evaluation of cancer cells, just like in the oral cavity tissue of patients. Therefore, the technique used for the isolation, culture, and evaluation of these cells is crucial.
The aim of the present study is to isolate and culture the cells from human primary Oral Squamous Cell Carcinoma [OSCC] tissue and evaluate them for morphological variations using an explant method.
The patients with OSCC who were undergoing surgery provided the tissue samples. An explant technique was used to achieve the isolation of cells from tissue samples. Following that, the cells were maintained, subcultured, and stored in accordance with the standard American Type Culture Collection [ATCC] protocol. Routine Hematoxylin & Eosin and crystal violet stains were used. These cells were morphologically studied, and the results were assessed for further studies.
We were able to successfully isolate and culture cells from 4 different tissue samples using the explant method. Morphological analysis revealed that one tissue had a significantly distinct presentation of epithelial and stromal cells, whereas the other three tissues had only minor morphological differences predominantly stromal cells. Two tissues were discarded after showing contamination.
Tissue culture should be done very meticulously specially when oral cavity tissue is used as it is house for millions of microorganisms. The technique must also be thoroughly followed and adjusted accordingly. Using common, inexpensive stains like Hematoxylin and Eosin and crystal violet, which are of great help for examining the morphology of cells routinely.
由于细胞培养在细胞生物学、生物技术和医学研究中的广泛应用,该技术已经发展成为一种广泛应用和接受的方法。从原发性癌症组织中分离原发性细胞是细胞培养技术的关键步骤,因为它为研究癌细胞的生物学、形态学和分子评估提供了可靠的来源,就像在患者的口腔组织中一样。因此,用于分离、培养和评估这些细胞的技术至关重要。
本研究旨在从人原发性口腔鳞状细胞癌 [OSCC] 组织中分离和培养细胞,并通过体外培养方法评估其形态学变化。
接受手术的 OSCC 患者提供了组织样本。使用组织块法实现从组织样本中分离细胞。然后,根据标准的美国典型培养物保藏中心 [ATCC] 方案保存、传代和储存细胞。常规使用苏木精和伊红以及结晶紫染色。对这些细胞进行形态学研究,并评估结果以进行进一步研究。
我们成功地使用组织块法从 4 个不同的组织样本中分离和培养细胞。形态学分析显示,一个组织的上皮细胞和基质细胞表现明显不同,而另外三个组织主要为基质细胞,形态差异较小。两个组织在显示污染后被丢弃。
特别是当使用口腔组织时,组织培养应该非常精细,因为它是数百万种微生物的家园。还必须彻底遵循技术,并进行相应的调整。使用苏木精和伊红以及结晶紫等常见且廉价的染色剂,对细胞形态进行常规检查非常有帮助。