Heterogeneity of the NIH3T3 Fibroblast Cell Line

Abstract

The embryonic mouse fibroblast cell line NIH3T3 is widely used in life science research, including the study of cell cycle control and primary cilia. Fibroblasts are the most important cell type in connective tissue, as they produce components of the extracellular matrix and determine tissue architecture. However, they are very heterogeneous and consist of subtypes specific to their organ of residence, among others. The NIH3T3 cell line was derived from whole mouse embryos that developed to pre-birth and is therefore most likely composed of different fibroblast subtypes. Furthermore, prolonged proliferation may have influenced their cellular composition. A heterogeneous cell population is unsuitable for any sophisticated research project. We found that the proportion of ciliated cells in the total NIH3T3 cell population was highly variable and asked whether this was a consequence of cellular heterogeneity and what molecular signatures were associated with it. We have established sub-cell lines by clonal expansion of single cells and characterized them morphologically and molecularly. Eventually, a myofibroblast-like and a fibroblast-like cell line were generated that differ in ciliation and proliferation. These homogeneous cell lines are valuable for a more detailed study of their molecular signatures, not least to uncover further the molecular pathways that contribute to the formation of the primary cilium.

ANKRD55 interacts with an IFT-B-like complex in microglia

“…Cilia formation in IMhu-A, IMhu-M, SH-SY5Y cells was probed using ARL13B Ab, a commonly used marker of primary cilia (43). NIH-3T3 cells, a prevalent model for the study of cilia, were used as positive control (44). Abs against IFT46 and IFT56 compatible with the rabbit ANKRD55 Abs used in this study are commercially not available; therefore, we settled on use of a compatible IFT81 Ab, and we validated some key findings with a goat IFT74 Ab in combination with a rabbit IFT46 Ab ( Supplementary Table 1 ).…”

TiB2-Derived Nanosheets Enhance the Tensile Strength and Controlled Drug Release of Biopolymeric Films Used in Wound Healing

“…Hence, the improved cell growth could also be attributed to the presence of boron moieties in the NTB films. The confocal image of cells on the 0.1% NTB film exhibited a “spindle-shaped” morphology, consistent with previous findings in the literature . Furthermore, there were no discernible variations in the cellular structure when compared to the control cells grown on a coverslip (Figure S12).…”

“…The confocal image of cells on the 0.1% NTB film exhibited a "spindle-shaped" morphology, consistent with previous findings in the literature. 94 Furthermore, there were no discernible variations in the cellular structure when compared to the control cells grown on a coverslip (Figure S12). Thus, from the results, it can be concluded that NTB films are biocompatible and promote cell adhesion without affecting the morphology of cells.…”

Encouraging tribomechanical and biological responses of hydroxyapatite coatings reinforced by various levels of niobium pentoxide particles

“…Overall, fibroblasts and osteoblasts are two types of cells that are in direct connection with the surface of the orthopedic implants. 73–76…”