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Frame et al. Investigating models for prostate cancer research

and so developing new treatments is an increasingly METHODS
complex task. Not only do we have to consider the
differences between patients, giving rise to the need Culturing of cell lines
and hope of patient stratification, but we also have to PNT1a, PNT2-C2 and LNCaP cells were cultured
consider that the tumor that has been biopsied may in Roswell Park Memorial Institute medium (RPMI)
be the larger more detectable one but not necessarily with 10% fetal calf serum. BPH-1 cells were cultured
[11]
the most aggressive one. Alongside that, we have our in RPMI with 5% fetal calf serum . PC3 cells were
dependence on hormone treatments for metastatic cultured in Hams-F12 media with 7% fetal calf serum.
prostate cancer, whilst knowing that there are tumor P4E6 cells were cultured in Keratinocyte Serum-Free
cell subpopulations that are not responsive or develop medium (KSFM) with supplements (bovine pituitary
acquired resistance to those treatments . There is a extract 50 mg/mL and human recombinant epidermal
[1]
[12]
poor choice of chemotherapy available for prostate growth factor 5 ng/mL) and with 2% fetal calf serum .
cancer and it is typically a last resort, although some To all media, Glutamine (2 mmol/L) was added. No
progress is being made in this area . Even with all antibiotics were used in the media. Cells were grown
[2]
o
these known variations, only in recent years has the in an incubator at 37 C in a humidified atmosphere
containing 5% CO .
true complexity of prostate cancers emerged [3-6] with 2
genomic and transcriptomic sequencing as well as Culturing of primary prostate cells
[7]
clonal tracking [8-10] . So, if we set out to test current Tumor tissue was obtained by targeted needle biopsy
treatments or develop novel therapies for prostate following radical prostatectomy. Following collection
cancer, we must consider our current drug pipeline from the hospital, tissue was digested overnight in
from bench to bedside; what models are used, do they collagenase, followed by a trypsin digest. Primary
take into account the different layers of heterogeneity prostate epithelial cells derived from patient tissue
and are they fit for purpose? were cultured in stem cell media (SCM). SCM
contains KSFM plus supplements (bovine pituitary
Here, we present a study that highlights the variation extract 50 mg/mL and human recombinant epidermal
in results that can be acquired when using different growth factor 5 ng/mL) with the addition of 2 ng/mL
cell line models, and also in comparison to primary stem cell factor, 100 ng/mL cholera toxin, 1 ng/mL
prostate epithelial cells cultured from patient tissue. granulocyte macrophage colony-stimulating factor and
We consider how to tackle the cellular heterogeneity 2 ng/mL leukemia inhibitory factor. Cells were cultured
within tumors by assessing cell subpopulations rather on Biocoat collagen I 10 cm dishes with irradiated
than a heterogeneous mixture, as well as introducing Sandoz inbred strain, thioguanine- and ouabain-
a new technique that might be instrumental in resistance (STO) feeder cells. A detailed method of
assessing drug response whilst simultaneously taking the whole procedure has been published . Patient
[13]
into account cell heterogeneity. samples used in this study are listed in Table 1.

Figure 1: Heterogeneity in prostate cancer. When considering the task of improving current prostate cancer treatments or developing novel
therapies, multiple types of heterogeneity have to be taken into account. These include patient tumor heterogeneity, multi-focal disease,
intra-tumor cellular heterogeneity, genomic heterogeneity including mutations and gene fusions and finally epigenetic heterogeneity with
inherent differences between cell populations but also the possibility of therapy-induced epigenetic changes
Journal of Cancer Metastasis and Treatment ¦ Volume 3 ¦ December 6, 2017 303