Abstract
Primary myeloma (PM) cells are short-lived in conventional culture, which limited their usefulness as a study model. Here, we evaluated if three-dimensional (3D) culture can significantly prolong the longevity of PM cells in-vitro. We employed a previously established 3D model for culture of bone marrow mononuclear cells isolated from 15 patients. We assessed the proportion of PM cells, viability and proliferation using CD38 staining, trypan blue exclusion assays and carboxy fluorescein succinimidyl ester (CFSE) staining, respectively. We observed significantly more CD38+ viable cells in 3D than in conventional culture (65% vs. 25%, p = 0.006) on day 3. CFSE staining showed no significant difference in cell proliferation between the two culture systems. Moreover, we found that PM cells in 3D culture are more STAT3 active by measure of pSTAT3 staining (66% vs. 10%, p = 0.008). Treatment of IL6, a STAT3 activator significantly increased CD38+ cell viability (41% to 68%, p = 0.021). In comparison, inhibition of STAT3 with Stattic significantly decreased PM cell viability in 3D culture (38% to 17% p = 0.010). Neither IL6 nor Stattic affected the PM cell viability in conventional culture. This study suggests that 3D culture can significantly improve the longevity of PM cells in-vitro, and STAT3 activation can further improve their viability.
Highlights
Immortalized human cancer cell lines have advanced cancer research since the 1950s, their suitability to represent real tumors has been challenged
We have previously shown that STAT3 signaling was activated in myeloma cell lines cultured in 3D [14]
We first tested if the 3D culture system we used previously can significantly increase the viability of Primary myeloma (PM) cells in-vitro, as compared to conventional culture
Summary
Immortalized human cancer cell lines have advanced cancer research since the 1950s, their suitability to represent real tumors has been challenged. To further complicate the issue, it was reported that 18% to 46% of human cancer cell lines are misidentified or cross-contaminated over time [1,2]. Several comparative studies between primary cells and cell lines had reported remarkable differences in their gene expression profiles [3,4,5]. A proteomic analysis comparing primary myeloma (PM) cells and myeloma cell lines showed significant differences in the expressions of biosynthesis proteins (such as ribosomal subunits, chaperons and translational factors) and immune response elements (such as complement receptors and MHC molecules) [7]. The sources for PM cells are relatively limited More important, it is well-known that PM cells, without appropriate support, remain viable in conventional culture for limited time (
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