ABSTRACT
Cell culture technology is the most commonly used method in the in vitro experiments at present. However, monolayer cell culture technology has been unable to meet the demand of the researchers. This is because that monolayer cell culture cannot mimic the cellular environment in which multiple cells interact with each other in the body. We cannot discuss the relationship of many cells, because we do not know the relationship between cells through a single kind of cell. So cell co-culture medicine arises at the historic moment for the demand. With the development of research method in recent years, cell co-culture method also has been improved in practice: from direct contact co-cultures to indirect contact co-cultures, from two-dimensional co-cultures to three-dimensional co-cultures. Cell co-culture method is closer to the human body. It is also more advantageous to study the interaction among cells. Nowadays, there are more researchers tend to select this method to study the physiological and pathological in vitro model, tissue engineering, and cell differentiation research. At the same time, it has become the focus of drug research and development, drug analysis, mechanism of drug action, and drug targets. This article will review the studies of cell co-culture method, summarize advantages and disadvantages of various methods, so as to promote improvement of cell culture methods, to build cells co-culture system that more close to human body, and build the in vitro model that simulate internal circulation of human body further.
ABSTRACT
OBJECTIVE: The aim of this study was to establish three-dimensionally cultured endometrial cell model containing endometrial stromal cell (ESC), endometrial epithelial cell (EEC) and extracellular matrix (ECM) and to compare the morphological and biomolecular expression patterns of this model with mid-luteal endometrium in vivo. MATERIALS AND METHODS: The EEC and ESC was obtained from hysterectomy specimen and cultured separately. The EEC was overlayered in Matrigel layer on ESC embedded in collagen. The model had been cultured for 48 h in DMEM medium containing estrogen and progesterone. The ultrastructure was evaluated by electron microscopy. The expression of integrins, cyclooxygenases and matrix metalloproteinases were examined by immunohistochemistry and zymography. RESULTS: EEC in three-dimensional culture model grew with polarity and tight junction and desmosome between cells were found. The formation of pinopodes was also detected. In three-dimensionally cultured endometrial cell model, the expression of integrin alpha1, alpha4, beta3, MMP-1, -2, -3 and 9 was detected which was not expressed in monolayer culture of EEC, ESC or ESC embedded in collagen. CONCLUSION: The three-dimensionally cultured endometrial cell model possessed the morphological and biomolecular characteristics of in vivo endometrium of implantation period. These characteristics could be achieved by paracrine interactions between ESC and EEC. This model may contribute to the studies of differentiation of endometrium, process of implantation and pathophysiology of implantation-related diseases.