三维微球旋转培养系统,3D spheroid Culture system,Stress-Free 3DTM cell culture system

型号:Stress-Free 3DTM cell culture system
联系人:李胜亮
联系电话:18618101725
品牌:celvivo

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
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
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

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

 BioArray Matrix 
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)

* The BAM is at the β-test version stage.