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celvivo三维旋转培养系统

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联系人:李胜亮
联系电话:18618101725
品牌:celvivo

3D Cell Culture Technology
Mimicking In vivo Performance

In vivo, cells in tissues and tumours 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

Why use 3D Spheroids

CELL FEATURE	Advantage over classical 2D cultures For HepG2-C3A - features for other cell lines maybe different
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)

Spheroid Characteristics

Spheroids

SPHEROIDS	Characteristics For HepG2-C3A - values for other cell lines will be different
Number of cells per spheroid (± 22%)	Day 8 = 23,156 Day 21 = 82,342 Media can be collected via one of the two ports
Spheroid size (Planimetric surface)	Day 8 = 0.26 mm? Day 21 = 0.58 mm?
Protein concentration	Day 8 = 21.71 ?g/ mm? Day 21 = 43.28 ?g/ mm?
ATP concentration	Day 8 = 9.23 ?M/ mm? Day 21 = 8.93 ?M/ mm?
Bioreactor soluble protein content	Day 8 = 2,010 ?g Day 21 = 10,120 ?g* * the culture will have been split into 4 bioreactors at this stage

Bioreactor

BIOREACTOR	Advantages
Easy access to samples and media	The bioreactor can be opened and closed again, providing a petri-dish like access to spheroids and media Repeated sampling possible Media can be collected via one of the two ports
Multiple samples	Bioreactor can cultivate up to 300 mature identical 'biopsies' which can either be collected at once for multiple assays or at multiple time points for fewer assays
Dynamic equilibrium	When non-lethal treatment stops, spheroids return to their resting state (dynamic equilibrium)
Optimal growth environment	Rotating bioreactor- clinostat system Inbuilt humidification chamber with good gas exchange Inert materials used: non-toxic and low unspecific binding
Use of different cell types	Cell from any species can be used Primary-, stem- and immortal cells have to be used in clinostat systems Co-culture of different cell types in the same bioreactor

* The Bioreactor patent is owned by MC2 Biotec (MC2biotec.com). Celvivo is commercialising it on their behalf.

BioArray Matrix Drive

Left (Video): Changing the growth media in the bioreactors and using the Bioarray matrix drive (BAM).
Right: One of the 16 proprietary drive axel speed controllers.

The drive's control board fits into a standard CO2 incubator (equipped with access port minimum of 24mm in diameter) and has 16 independently control drive axels which can rotate in either direction at speeds from 5 to 50 rpm. The system is temperature inert for the incubator. It can be controlled either from windows or android based devices.

BIOARRAY MATRIX DRIVE	Advantages
Optimal growth environment for 3D culture	Rotating bioreactor- clinostat Humidified chamber with good gas exchange Inert materials used: non-toxic and low unspecific binding Computer controlled multiple drive unit Does not disturb incubator temperature
Multiple bioreactors	16 independent drives 5-50 rpm in
Easy control	Bioreactor rotation speed can be regulated from – a tablet (great for working in the cell culture lab) – a network computer – or smart phone (using the app provided) Drives can be regulated independently or in any combination of groups
Easy overview	Large simple display on tablet Internal log
Quick suspension option	Special start-up program to rapidly disperse spheroids after bioreactor has been stopped (e.g. for medium change or treatment)