In Vitro Efficient Expansion of Tumor Cells Deriving from Different Types of Human Tumor Samples

Ilaria Turin,Barbara Bello,Arsenio Spinillo,Daniela Montagna,Marcello Maestri,Ombretta Luinetti,Marco Paulli,Marianna Roccio,Paolo Dionigi,Matteo Tanzi,Paolo Pedrazzoli,Rita Maccario,Federica Ferulli,Roberta Schiavo

doi:10.3390/medsci2020070

Abstract

Obtaining human tumor cell lines from fresh tumors is essential to advance our understanding of antitumor immune surveillance mechanisms and to develop new ex vivo strategies to generate an efficient anti-tumor response. The present study delineates a simple and rapid method for efficiently establishing primary cultures starting from tumor samples of different types, while maintaining the immuno-histochemical characteristics of the original tumor. We compared two different strategies to disaggregate tumor specimens. After short or long term in vitro expansion, cells analyzed for the presence of malignant cells demonstrated their neoplastic origin. Considering that tumor cells may be isolated in a closed system with high efficiency, we propose this methodology for the ex vivo expansion of tumor cells to be used to evaluate suitable new drugs or to generate tumor-specific cytotoxic T lymphocytes or vaccines.

Highlights

Tumor cell lines represent a unique tool for investigating both tumor biology/profiles and the mechanisms underlying tumor responsiveness to anticancer therapies [1,2,3]
All primary cultures were maintained for at least three weeks in culture before phenotypical analysis and a portion of early passage cultures was frozen for further studies
Cultures from metastases were generally more difficult to maintain in culture and we only reached a maximum of 11 passages, while cultures from primary tumors generally grew well and were maintained for 24–30 passages

Summary

Introduction

Tumor cell lines represent a unique tool for investigating both tumor biology/profiles and the mechanisms underlying tumor responsiveness to anticancer therapies [1,2,3]. The establishment of tumor cell lines is extremely difficult and the success rate is low and unpredictable [4]. Tumors exhibit apparent autonomy from normal regulatory control in vivo, they often fail to grow when cultured in vitro [2]. Other than neoplastic cells, may be obtained from tumor samples, including connective-tissue fibroblasts, infiltrating lymphocytes and elements of normal tissue from which the neoplasia arose. When trying to establish a tumor cell line, the major problem is contamination by fibroblasts or other types of endothelial cells, which grow readily in culture and may respond to tumor-derived mitogenic factors [5]. In primary tumor cultures many cells may not be capable of propagation due to genetic or phenotypic aberrations, terminal differentiation or nutritional insufficiency. Some primary cultures can be subcultured, opening up major research possibilities [6,7]

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