Why use 3D? 

In vivo, cells in tissues and tumors do not double every 2 – 4 days as they do in classical (2D) cell culture – so why use a technology developed in the 1950’s? 

The CelVivo system creates an environment which promotes the growth and maintenance of large 3D tissue mimetic structures, whether they are spheroids, organoids, acini and other aggregates. 

In this environment, liver spheroids remain in a stable dynamic equilibrium and exhibit human in vivo physiological performance for at least 24 days (i.e. up to at least 42 days of culture). During this time, the spheroids exhibit a stable transcriptome profile. 

Reasons for Using 3D Cell Culture

Proliferation

Slows from a doubling time of 1 day to 60 days after 40 days in culture (see reference 3)
Resembles proliferation in the parental tissue or in tumours in vivo
Cultures can be maintained for over 300 days 

Architecture

Microfilaments (actin) microtubules (tubulin) and intermediate filaments (e.g. keratin) resemble that seen in vivo(see reference 7)

Ultrastructure

Resembles liver sections, showing plasma membrane differentiation, tight junctions, bile canaliculus-like channels, lipid droplets and glycogen granules (see reference 2) 

Physiological performance

Levels of ATP, urea and cholesterol reach levels seen in vivo (see reference 3)

Drug metabolism

Toxicity measurements for Amiodorone, APAP, Metformin, Phenformin and Valproic acid are more accurate than those obtained using human primary hepatocytes (see reference 1)

Proteomics

Numerous changes can be detected throughout the proteome (see reference 7)

Metabolic reprogramming

> 21 day old spheroids show clear aerobic metabolism (Warburg effect) (see reference 7)

Epigenetics

Spheroids display histone marks and clipping (not seen using 2D cultures) (see reference 9)

Integration of cellular response

APAP causes protein oxidation and nitrosylation and this integrates the cellular response to physiological doses of APAP (see reference 11)