Establishment of A Three-Dimensional Culture Condition for The U266 Cell Line Based on Peripheral Blood Plasma-Derived Fibrin Gels

Document Type : Original Article


1 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Applied Cell Sciences Division, Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran


Objective: The study of pathophysiology as well as cellular and molecular aspects of diseases, especially cancer,
requires appropriate disease models. In vitro three-dimensional (3D) structures attracted more attention to recapitulate
diseases rather than in vitro two-dimensional (2D) cell culture conditions because they generated more similar
physiological and structural properties. Accordingly, in the case of multiple myeloma (MM), the generation of 3D
structures has attracted a lot of attention. However, the availability and cost of most of these structures can restrict their
use. Therefore, in this study, we aimed to generate an affordable and suitable 3D culture condition for the U266 MM
cell line.
Materials and Methods: In this experimental study, peripheral blood-derived plasma was used to generate fibrin gels
for the culture of U266 cells. Moreover, different factors affecting the formation and stability of gels were evaluated.
Furthermore, the proliferation rate and cell distribution of cultured U266 cells in fibrin gels were assessed.
Results: The optimal calcium chloride and tranexamic acid concentrations were 1 mg/ml and 5 mg/ml for gel formation
and stability, respectively. Moreover, the usage of frozen plasma samples did not significantly affect gel formation and
stability, which makes it possible to generate reproducible and available culture conditions. Furthermore, U266 cells
could distribute and proliferate inside the gel.
Conclusion: This available and simple fibrin gel-based 3D structure can be used for the culture of U266 MM cells in a
condition similar to the disease microenvironment.


Main Subjects

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