Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids

Author(s)

Bonazza, Camila ; Andrade, Sheila Siqueira ; Sumikawa, Joana Tomomi ; Batista, Fabricio Pereira ; Paredes-Gamero, Edgar J. ; Girao, Manoel J. B. C. ; Oliva, Maria Luiza V. ; Castro, Rodrigo Aquino

• Document type: Journal article
• Source: PLoS One; v. 11, n. 7 JUL 8 2016.
• Web of Science Citations: 1

Abstract

Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 beta-estradiol (E-2) and progesterone (P-4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity. (AU)

• FAPESP's process: 12/19851-8 - Heterotypic culture: The Cross-Talking between epithelial tumor cells, mammary stroma, macrophages and platelets in breast cancer: the release of proteolytic enzymes and signaling of Toll-like receptors
• Grantee: Sheila Siqueira Andrade
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 09/53766-5 - Proteins from plant source with selectivity for inhibition of mammalian enzymes and their role as an anti-inflammatory, antithrombotic, anti-diabetic and anti-tumor agent
• Grantee: Maria Luiza Vilela Oliva
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/19780-3 - Heterotypic culture: the cross-talking between epithelial tumor cells, mammary stroma, macrophages and platelets in breast cancer: the release of proteolytic enzymes and signaling of toll-like receptors
• Grantee: Manoel João Batista Castello Girão
• Support Opportunities: Regular Research Grants