Extracellular matrix components cell culture

Cell culture studies have shown that aldehydic products derived from ethanol metabolism and lipid peroxidation can increase collagen mRNA levels and enhance the expression of connective tissue proteins. Acetaldehyde is able to increase the production of several extracellular matrix components. Studies also show that hepatic stellate cells, which are the primary source of extracellular matrix, become readily activated under conditions involving enhanced oxidative stress and lipid peroxidation. 

While tissue isolation, immortalization, and proliferative status all can severely impact cell culture differentiation, other parameters are also critical such as nutrient concentrations, serum content, hormones, growth factors and cytokines, culture substrata, extracellular matrix components, and oxygen tension. 

For the cultivation of airway epithelial cells, the cells are usually plated onto tissue culture plates coated with collagen or extracellular matrix components. Hormonally defined Ham s F12 medium contained 1 % penicillin-streptomycin, 5 ig/ml insulin, 5 ig/ml transferrin, 25 ng/ml epidermal growth factor, 15 ig/ml epithehal growth factor supplement, 2 X 10" M triiodothyronin, and 10" M hydrocortisone. 

Fukuda, J., A. Khademhosseini et al. 2006. Micropatterned cell co-cultures using layer-by-layer deposition of extracellular matrix components. Biomaterials 27(8) 1479-1486. 

Additional cell culture experiments were carried out on patterned 3D substrates in order to combine the spatial constraints of a stem cell niche with the influence of different extracellular matrix components of its microenvironment. Therefore, sihcone molds of cavities with a size of 10 to 80 p,m and a depth of 10 pim were coated with reactive maleic anhydride copolymers and subsequently reconstituted assembUes of collagen fibrils and cofibrils were immobilized on top of it. Hematopoietic stem cells were seeded onto these functionalized cell carriers. Depending on the size of the cavities cells differentially adhered to these structmes as exemplarily shown in the inset of Fig. 10. In initial experiments adhesion and proliferation was followed over a time period of 3 days. As shown in Fig. 10 the cells tend to preferentially adhere to an intermediate size of cavities. This result points to a balanced equilibrium of cell-ceU contacts and cell-matrix contacts for the homing and proUferation of hematopoietic stem cells, which is in agreement with the observed small hematopoietic stem cell clnsters foimd in vivo [113]. Fm-ther experiments will foUow np on these observations in order to imravel the influence of different extracellular matrix compositions combined with microcavities. 

F9 embryonal carcinoma cells have a simple set of growth supplements which are required for growth in serum-free medium insulin, transferrin, and fibronectin (Rizzino and Sato, 1978). Fibronectin is a component of the extracellular matrix and facilitates the attachment of the cells to the culture dish. In addition, high density lipoprotein (HDL) has been observed to promote the growth of F9 cells serum-free. 

In vitro and in vivo studies have shown that stromal cells provide a rich environment of signals (cytokines, extracellular matrix, adhesion molecules etc.) that control proliferation, survival and differentiation of hematopoietic progenitor cells (Chailakhian, 1978 Verfaillie, 2002). Recently investigators incorporated stromal components into the expansion cultures (Rawano et al. 

There are complex interactions among the different cellular components of the neurovascular unit and the extracellular matrix, determining its permeability properties during both physiological and pathological conditions. This highlights the severe limitations of cell culture-based models to mimic neurological diseases associated with BBB disruption. Transwell culture systems of endothelial cells alone rarely achieve adequate transendothelial electrical resistance (TEER). Cocultures of... 

The development of parallel-plate perfusion chambers [67,68] made possible the study of platelet interaction with the extracellular matrix (ECM) generated by cells in culture or with isolated subendothelial components under defined experimental conditions. The use of the ECM produced by human umbilical vein endothelial cells (HUVEC) in culture as adhesive substrate has Su tated the understanding of the mechanisms involved in primary hemostasis [68]. HUVECs are immature and not subjected to flow conditions during their culture, two Ikctors which may influence the reactivity of their ECM towards platelets [69]. Interestingly, the properties and reactivity of the underlying ECM can be modified by exposure of HUVECs to different stimuli, an experimental approach which has fevored the investigation of basic mechanisms of thrombosis [33]. 

Owing to their stmctural similarity to the macromolecular-based components in the body (and their biocompatibility, permeability, and physical characteristics), hydrogels can also serve as a synthetic extracellular matrix (ECM) to organize cultured cells into a three-dimensional architecture and to present stimuli that direct the growth and formation of a desired tissue [33]. The balance between mechanical properties and controlled degradation of hydrogels mainly depends on the original... 

Most cultured vertebrate cells will grow only when attached to a negatively charged substratum that Is coated with components of the extracellular matrix. 

Although the system contemplates the distinction of chemotaxis and chemokinesis, it does not account for the complexities of the extravasation process. To emulate these, the filter can be coated in advance. Even though some studies use components of the extracellular matrix such as collagen, we have found that the culture of endothelial cells on top of the filter optimally mimics in vivo conditions. Similarly, other chemotaxis assays are available in the market, most of them based on a two-chamber model. For instance, the Zigmond chamber and its improvement, the Dunn chamber allow for the visuahzation of the cell migration toward the chemoattractant, but they do not allow for a more in vivo simulation (Sackmann, Fulton, Beebe, 2014). 

The development of a cellular microenvironment in a microfluidic chip starts with device fabrication. The most commonly used material for fabrication is polydimethylsiloxane (PDMS). One of the most important components to develop such platforms is the extracellular matrix (ECM), which is the 3D cellular microenvironment. Different approaches have been followed to pattern the ECM inside the microfluidic device, which will significantly affect the arrangement of cells on the chip. Following this, the target cells are seeded and cultured. Such cell culture techniques would be common for drug screening as well as the fundamental research. But for CTCs detection, usually the antibodies will be immobilized on the chip before introducing the cells. 

---