Author : tempo

Written on Jul, 08, 2020 by in ,
TempoATP – a snapshot

Genetically encoded biosensors have become popular and powerful LIVE-cell reporters in recent years (see reference here). These biosensors can be incorporated into a variety of human inducible pluripotent stem cells (iPSCs) and iPSC-derived cell types (such as neurons, glial cells, kidney cells, and cardiomyocytes, just to name a few). Previously, we discussed the biosensors and how they are used in research (here). TempoATP biosensor (image shown above) is a LIVE-cell reporter that tracks intracellular ATP levels in real time (from seconds to hours).

Examples of where mitochondrial ATP productions are important?

#1: Research from neurodegenerative disorders (e.g., Alzheimer’s, amyotrophic lateral sclerosis (ALS), Huntington’s, and Parkinson’s) has shown the importance of mitochondria in neuronal survival. In other words, ageing and neurodegeneration depend on mitochondrial health (see reference here).

#2: Cancer research has shown that mitochondrial production of ATP is associated with calcium homeostasis. Furthermore, tumorigenesis depends on ATP production and mitochondrial function (see reference here).

#3: Mitochondrial dysfunction and oxidative stress are key characteristics found in the ageing human skin (see reference here). Some of the notable phenotypes are: wrinkle formation, hair loss, uneven pigmentation, inflammation, photo-aging, and wound healing impairment (see reference here).

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Written on Jun, 09, 2020 by in , ,

Spheroids have been used in cell culture for decades. In the 1980s, different types of human cancer cells –normally grown as monolayers or suspension cultures–were tested for their innate abilities to form and grow as spheroids in vitro (1). 16 out of 27 tested tumor cell lines successfully formed spheroids (1). In the tumor cells study, scientists also observed that all large spheroids had necrotic centers but their shapes varied. Another study used V79 hamster cells (V79 379A), a human small cell carcinoma of the lung (ME/MAR) and 2 xenographed human melanomas (HX117 and HX118) in spheroids cultures in the 1980s (2). In the study, the effects on spheroid growth due to radiation treatments were measured and evaluated (2).

Widely acknowledged is the unique ability of spheroids to mimic natural cell responses and interactions. Cells in 3D are more representative of their native conditions than the traditional 2D monolayer culturing conditions (e.g., cell-to-cell interactions, drug-induced responses, and cells-to-environment responses). Furthermore, the multicellular arrangement allows different cell types to interact with each other within each spheroid. Past studies examined growth rates, hypoxic conditions, and other survival conditions for the spheroids (1-2). Currently, many cell culture reagents and instruments are available to enable spheroid cultures, making the technique increasingly accessible, flexible and approachable for scientists in a variety of research fields. (Unlock your creative minds!)

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Written on May, 01, 2020 by in ,

Vascular endothelium is a thin monolayer of cells that constitute the lining of blood vessels and organs (1-3). A hallmark of many diseases (e.g., cancer, diabetes mellitus, viral infections, etc) is highlighted in the dysfunctional states of the vascular endothelium. Vascular endothelial cells line the entire circulatory system — heart, large blood vessels, and small capillaries (1). And the cells form the barrier between blood and organ tissues (3). 

Notable functions of the vascular endothelium:

1) Controls and regulates vascular relaxation and constriction;

2) Regulates homeostasis of solutes, fluid, macromolecules, hormones, platelets, and blood cells;

3) Directs “foreign materials” to inflammatory cell types;

4) Regulates blood fluidity;

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