Expansion of airway basal epithelial cells from primary human non‐small cell lung cancer tumors

Abstract

Pre‐clinical non‐small cell lung cancer (NSCLC) models are poorly representative of the considerable inter‐ and intra‐tumor heterogeneity of the disease in patients. Primary cell‐based in vitro models of NSCLC are therefore desirable for novel therapy development and personalized cancer medicine. Methods have been described to generate rapidly proliferating epithelial cell cultures from multiple human epithelia using 3T3‐J2 feeder cell culture in the presence of Y‐27632, a RHO‐associated protein kinase (ROCK) inhibitor, in what are known as “conditional reprograming conditions” (CRC) or 3T3 + Y. In some cancer studies, variations of this methodology have allowed primary tumor cell expansion across a number of cancer types but other studies have demonstrated the preferential expansion of normal epithelial cells from tumors in such conditions. Here, we report our experience regarding the derivation of primary NSCLC cell cultures from 12 lung adenocarcinoma patients enrolled in the Tracking Cancer Evolution through Therapy (TRACERx) clinical study and discuss these in the context of improving the success rate for in vitro cultivation of cells from NSCLC tumors.

Conditional reprogramming: next generation cell culture

“…The irradiated J2 cells can directly serve as feeders. Other way is to use the anti-proliferative mitomycin C (2–4 μg/mL) to treat J2 cells for 1–3 h to acquire feeder cells 25 . However, it is necessary to wash the mitomycin C-treated cells for several times to avoid growth inhibition by mitomycin C during co-cultures.…”

“…The eligibility of CR to immortalize a wide range of epithelial cells of varied origins has been proved by many researches. Primary cells from both normal and pathological tissues can be proliferated under CR conditions ( Table 2 12 , 25 , 69 , 83 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 , 126 , 127 , 128 , 129 , 130 , 131 , 132 , 133 , 134 , 135 , 136 , 137 , 138 , 139 , 140 , 141 , 142 , 143 , 144 , 145 ). Of particular note, not only human tissues but also tissues from other origins such as mouse, horse, dog and fish have been used for CR culture.…”

“…In the study of Hynds et al. 25 , CR condition only enabled tumor cell culture from 1 out of 10 primary NSCLC tumors. They suggested that expanded CR tumor cells was seen at passage 2, which was rapidly outgrown by normal epithelial cells in later passages 25 .…”

“… 25 , CR condition only enabled tumor cell culture from 1 out of 10 primary NSCLC tumors. They suggested that expanded CR tumor cells was seen at passage 2, which was rapidly outgrown by normal epithelial cells in later passages 25 . Notably, this CR tumor cells could form a tumor in NSG mice and re-culture of cells from the tumor xenograft maintained key characteristics of primary tumor 25 .…”

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.…”