Receptor Tyrosine Kinase Targeting in Multicellular Spheroids

Abstract

While growing cells as a monolayer is the traditional method for cell culture, the incorporation of multicellular spheroids into experimental design is becoming increasingly popular. This is due to the understanding that cells grown as spheroids tend to replicate the in vivo situation more reliably than monolayer cells. Thus, the use of multicellular spheroids may be more clinically relevant than monolayer cell cultures. Here, we describe methods for multicellular 3D spheroid generation that may be used to provide samples for receptor tyrosine kinase (and other protein) detection. Methods described include the forced-fl oating poly-HEMA method, the hanging-drop method, and the use of ECM to form multicellular 3D spheroids.

Tyrosine kinase inhibitors target cancer stem cells in renal cell cancer

“…3D cell culture. In the 3D approach, CSCs were grown in media as above but as aggregates and in hanging drop culture forming spheroids as previously described (44,45), with modification of 100 cells/10 µl drop. After formation of the spheroids (72 h), sunitinib was added in a volume of 5 µl to each drop in order to obtain 0, 1, 2, 4, 8, and 15 µM concentrations.…”

Levitational 3D Bioassembly and Density‐Based Spatial Coding of Levitoids

“…However, bi‐dimensional culture is far from representing an accurate mimicry to in vivo microenvironment due factors such as lack of proper cell–cell and cell–ECM interactions, [ 7 ] oxygen and nutrient distribution patterns, [ 8 ] and morphological characteristics. [ 9 ] Taken together, a broad practical application to generate 3D soft materials out of living cells is still needed to bridge the gap between monolayer cell culture and in vivo. [ 10 ]…”