Effective method for the isolation and proliferation of primary lung cancer cells from patient lung tissues

Abstract

We have developed a technique for isolating and culturing primary lung cancer cells extracted from patient tissue to facilitate anti-cancer drug development. Patient-derived lung cancer tissues were mechanically dissociated to 40-100 μm. Dispase was then used to isolate cultured lung cancer cell populations, which were re-plated on Matrigel-coated dishes containing N2-supplemented medium and growth factors. This method allows pure populations of primary non-small cell lung cancer cells to be grown in vitro. The isolated cells exhibited hallmark cancerous properties such as abnormal chromosomes and in vivo tumor formation. The cell lines generated through this procedure may help to advance our knowledge of certain forms of lung cancer and may also be useful for developing patient-specific anti-cancer drug screening procedures.

Summary of Primary Culture of Human Lung Cancer Cells

“…A study indicates [4] that media containing serum or growth factors promote the growth of fibroblasts, making it difficult to purify the required cell population in the co-culture state. H. Baillie-Johnson et al [6] cultured Small Cell Lung Cancer (SCLC) cell lines and found that when these fibroblasts began to overgrow, to transfer specimens growing in HITES+2.5% FCS to separate HITES alone for some period of time could inhibit fibroblast growth.…”

“…PingWang et al [7] found that the culture medium of Defined Keratinocyte-Serum Free Medium (DK-SFM) did support the selective growth of typical epithelial cells and significantly inhibited the growth of fibroblasts coexisting. Joseph Seo et al [4] indicated that replacing serum with N2 supplements prevented fibroblast proliferation while allowing lung cancer cells to grow. As a result, the N2-supplemented was superior in purification while the BEI medium of 10% FBS was the first-rank culture medium in proliferation.…”

“…The use of other enzymes: Joseph Seo [4] raised the problem of using enzymes to isolate lung cancer cells. They used many enzymes, such as collagenase, disperse enzyme, trypsin, and so on, but the results were not satisfactory, because it was difficult to get a single cell population after enzyme treatment.…”

“…Joseph Seo et al [4] pointed out tumor staging affects the expression of EMT (Epithelial-Mesenchymal Transition) markers.…”

Recent Approaches Encompassing the Phenotypic Cell Heterogeneity for Anticancer Drug Efficacy Evaluation

“…The first attempt to solve the disadvantage of cell line "purity" as well as personalization of the treatment approach was the introduction of primary ex vivo cancer cell lines into the cancer research, including drug testing, discovery and therapy response prediction, with improving methodologies and optimizations for cell culturing [175][176][177][178]. Heterogeneity found in cell cultures was suggested to be coded by biological mechanisms from the primary tumors, and it was acknowledged as an instrument for clinical implications.…”