A novel human leiomyoma tissue derived matrix for cell culture studies

Abstract

BackgroundThe composition of the matrix molecules is important in in vitro cell culture experiments of e.g. human cancer invasion and vessel formation. Currently, the mouse Engelbreth-Holm-Swarm (EHS) sarcoma -derived products, such as Matrigel®, are the most commonly used tumor microenvironment (TME) mimicking matrices for experimental studies. However, since Matrigel® is non-human in origin, its molecular composition does not accurately simulate human TME. We have previously described a solid 3D organotypic myoma disc invasion assay, which is derived from human uterus benign leiomyoma tumor. Here, we describe the preparation and analyses of a processed, gelatinous leiomyoma matrix, named Myogel.MethodsA total protein extract, Myogel, was formulated from myoma. The protein contents of Myogel were characterized and its composition and properties compared with a commercial mouse Matrigel®. Myogel was tested and compared to Matrigel® in human cell adhesion, migration, invasion, colony formation, spheroid culture and vessel formation experiments, as well as in a 3D hanging drop video image analysis.ResultsWe demonstrated that only 34 % of Myogel’s molecular content was similar to Matrigel®. All test results showed that Myogel was comparable with Matrigel®, and when mixed with low-melting agarose (Myogel-LMA) it was superior to Matrigel® in in vitro Transwell® invasion and capillary formation assays.ConclusionsIn conclusion, we have developed a novel Myogel TME matrix, which is recommended for in vitro human cell culture experiments since it closely mimics the human tumor microenvironment of solid cancers.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1944-z) contains supplementary material, which is available to authorized users.

Interface between breast cancer cells and the tumor microenvironment using platelet-rich plasma to promote tumor angiogenesis - influence of platelets and fibrin bundles on the behavior of breast tumor cells

“…CD95 L plays an important tumor supportive role in both breast tumor cells and endothelial cells controlling the angiogenic process. CD95 L (soluble), or Fas-L is an effector regulator of endothelial cell survival in recently formed endothelial cells related with the cytokines identified in our screen, including IL-8 and VEGF [42, 43]. Using an antibody specific to the CD95 L soluble form, we observed that the levels of CD95 L in the flow cytometry analysis were negligible in the FBS and PPP supplemented control breast tumors cells (data not shown).…”

Human Tumor Tissue-Based 3D In Vitro Invasion Assays

“…Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells are the prevailing substrates in the majority of in vitro cell invasion studies [1]. We have recently described a novel protein extract made of human uterus leiomyoma tissue and demonstrated that it is superior to Matrigel in in vitro cancer and angiogenesis studies [2]. However, such protein mixtures lack the complex 3D architecture which also influences proteolytic actions during cell invasion [3].…”

3D Culture Histology Cryosectioned Well Insert Technology Preserves the Structural Relationship between Cells and Biomaterials for Time‐Lapse Analysis of 3D Cultures

“…These collagen‐containing matrices may well be lacking collagen fiber structures required for birefringence . Matrigel and Myogel are mechanically and chemically processed preparations . As for the cell clusters in Matrigel, inadequate alignment of the newly synthesized immature collagen fibers is not causing the positive signal similar to mature collagen fibers.…”

“…For the liquid biomaterials in the 3D‐cryo well inserts we placed cells in Myogel (a human leiomyoma‐derived matrix suspension in media) and in cell culture media. The preparation of the biomaterial Myogel has been described in detail in another publication . The Ethics Committee of Oulu University Hospital approved our use of the myoma tissue obtained from a patient's uterus leiomyoma.…”

“…Other groups have attempted to engineer salivary structures in biomaterials, including hyaluronic acid, polyethylene glycol, Matrigel, collagen gels, placenta basement membrane extract, and fibronectin . To test the applicability of the 3D‐cryo well insert, we tested it in four soft biomaterials: Matrigel, GrowDex, Myogel, and a composite Myogel + GrowDex.…”