Culturing Uveal Melanoma Cells

Abstract

A major challenge in cancer research is the use of appropriate models with which to study a specific biological question. Cell lines have long been used to study cellular processes and the effects of individual molecules because they are easy to use, grow rapidly, produce reproducible results and have a strong track record in research. In uveal melanoma in particular, the absence of animal models that faithfully replicate the behavior of the human disease has propagated the generation and use of numerous cell lines by individual research groups. This in itself, however, can be viewed as a problem due to the lack of standardization when characterizing these entities to determine how closely they reflect the genetic and phenotypic characteristics of this disease. The alternative is to use in vitro primary cultures of cells obtained directly from uveal melanoma patient samples, but this too has its difficulties. Primary cell cultures are difficult to use, hard to obtain and can show considerable heterogeneity. In this article, we review the following: (1) the uveal melanoma cell lines that are currently available, discussing the importance of establishing a bank of those that represent the molecular heterogeneity of uveal melanoma; (2) the methods used to isolate and perform short-term cultures of primary uveal melanoma cells, and (3) the establishment of 3D tissue culture models that bridge the gap between 2D in vitro systems and in vivo models with which to dissect cancer biology and perform therapeutic screens.

Characterization of luteinizing hormone-releasing hormone receptor type I (LH-RH-I) as a potential molecular target in OCM-1 and OCM-3 human uveal melanoma cell lines

“… 20 , 21 , 38 – 44 In UM research, cell lines have been widely used in order to identify and characterize the disease in vitro. 45 In the present study, our aim was to investigate the mRNA expression for LH-RH ligand and LH-RH-I receptors in OCM-1 and OCM-3 human UM cell lines. Those cultures were derived from primary UMs.…”

Novel mouse model for primary uveal melanoma: a pilot study

“…This would be an ideal uveal melanoma model of primary uveal melanoma as well as metastasis formation. However, concerns regarding the cell line origin of OCM8 have been raised on the basis of the detection of a BRAF mutation that is not usually found in uveal melanoma cells . Yang et al injected Mel290 human uveal melanoma cells into the posterior compartment of the eyes of immunodeficient NU/NU mice and detected tumour growth in the eye as well as the formation of liver micrometastasis in 100% of the animals.…”

“…However, concerns regarding the cell line origin of OCM8 have been raised on the basis of the detection of a BRAF mutation that is not usually found in uveal melanoma cells. 44 Yang et al injected Mel290 human uveal melanoma cells into the posterior compartment of the eyes of immunodeficient NU/NU mice and detected tumour growth in the eye as well as the formation of liver micrometastasis in 100% of the animals. In particular, Mel290 uveal melanoma cells do not have GNAQ or GNA11 mutations and do not express any melanocyte-associated lineage markers that differentiate these cells from most other uveal melanoma cells.…”

“…In particular, Mel290 uveal melanoma cells do not have GNAQ or GNA11 mutations and do not express any melanocyte-associated lineage markers that differentiate these cells from most other uveal melanoma cells. 44 In contrast, our model uses human-derived uveal melanoma cells (UMT2) that express many different melanocyte-associated lineage markers and demonstrate a mutation in GNA11, as do many uveal melanomas. 17 Our mouse model with the UMT2 cell line allows the investigation of tumour biology of primary uveal melanoma because of the high similarity between the tumours generated in the mouse eyes and uveal melanoma in human eyes.…”

CD166high Uveal Melanoma Cells Represent a Subpopulation With Enhanced Migratory Capacity

“…Culture of PUM cells was also performed as previously published. 15 The fresh tumor tissue was minced using a sterile blade. The tissues pieces were then resuspended in collagenase IV (Sigma Aldrich Corp.) and incubated at 378C for 1 hour.…”