CORNING Life Sciences

Cell Culture Treatments

Corning A brief History

- A brief history
- Cornings Brands
- Cornings Workflows
-

Cell Culture Workflow

- Cell Culture surfaces

-

Suspension cells
- Non treated
- ULA = Ultra Low Attachment

Adherent Cells
-

TC = standard treated culture

Synthetic non specific
Biological coating
Syntethic defined
Syntehtic specific

- Other products with specific cell culture surfaces

-

Inserts cell culture

Assays microplates
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Corning A brief History

Founded in 1851 : world leader in specialty glass and ceramics.
Corning created and make keystone components that enable high-technology
systems for consumer electronics, mobile emissions control, telecommunications
and life sciences : bubble (first one it Edison ; touchable screens ;
Headquarters : Corning, New York
Employees:
Approximately 25,000 worldwide
Corning acquired Costar Corporation to become Corning Costar in 1993.
Corning acquired Axygen end of 2009
Corning acquired Gosselin end of 2010
Corning acquired Falcon DL end of 2012

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Corning Life Sciences Different Brands & Market segments

Whatever is your application, you can now get a more comprehensive line of lab
research tools with Cornings quality.

FALCON

Cell Culture, Drug Discovery &

Liquid Handling

Molecular Biology
Genomics

Microbiology

cellgro is available in Europe as Mediatech.

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Corning Life Science - Workflows

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Corning Life Science - Workflows

Cell Culture surfaces :
- Suspension cells
- Adherent cells
Adherent Cells :
- Standard - TC
- Synthetic improved
(CellBIND ; Primaria)

- Biological
(ECM Biocoat)

- Synthetic defined
(Purecoat)

- Synthetic specific
(OsteoAssays / Synthemax)

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Corning Life Science Cell Culture surfaces

Adherent culture
Moving
Roller
bottles
Microcarriers
in spinner
flasks
Flasks on
rockers

Stationary
Flasks
Microplates
Dishes
Transwell
inserts
HYPERFlask
CellSTACK

Suspension culture

Moving
Spinner
flasks
Shaker
flasks
Roller
bottles
Bags

Stationary
Flasks
Storage
bottles
Dishes
Microplates
Transwell
inserts
Bags
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Corning Life Science Cell Culture surfaces

Adherent culture

Suspension culture

Moving

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Corning Life Science Cell Culture surfaces

Suspension Cells
Suspension Cell Culture :
NT (Not Treated)
For standard suspension cell culture

ULA (Ultra Low Attachment)

For advanced suspension cell applications (stem cells..)

2013 Corning Incorporated

Corning Life Science Cell Culture surfaces

Adherent Cells
Adherent Cell Culture :

Animal Free

Specificity to Cells

YES

- Standard TC
- Synthetic improved (CellBIND ; Primaria)

YES

- Biological (ECM Biocoat)

NO

- Synthetic defined (Purecoat)

YES

- Synthetic specific (OsteoAssays / Synthemax)

YES

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Cell Culture surfaces - Adherent Cells

Standard Tissue Culture Surfaces
Standard Tissue Culture (TC) Treatement :
To be used with adherent cells, the Polystyrene needs to be treated to
add charges and to get it hydrophylic.
Polystyrene is fully hydrophobic and no natural.
TC-surface treatment is achieved by (similar proceedure) :
By gas plasma : placing polystyrene vessels into a vacuum chamber that
receives a mixture of gases with electric power.
By corona discharge.

The treatment will create a modification of the polystyrene altering its

surface chemistry Negative charges
Feature No specific
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Cell Culture surfaces - Adherent Cells

Standard Tissue Culture Surfaces

Culture
Vessels

- uncharged
- hydrophobic
- No cell attachment

Corona Discharge Plate

High energy oxygen ions

Conveyor belt

e.g. "non-treated"

rounder

40 ml droplet

Surface
treatment

H C

C
H

C
H

OH H

HO

Hydrophilic

OH

Hydrophobic

- charged groups
- hydrophilic
- cell attachment

O
C
OH

flatter

e.g. "TC"

40 ml droplet

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Cell Culture surfaces - Adherent Cells

Synthetic Non Specific CellBIND & Primaria
Syntethic Non specific (Improved attachment) CellBIND or Primaria :
To be used with adherent cells, the Polystyrene needs to be treated to
add charges and to get it hydrophylic.
Polystyrene is fully hydrophobic and no natural.
TC-surface treatment is achieved by :
a specific gaz mixture.
A difference source of energy (microwaves).

The treatment will create a modification of the polystyrene altering its

surface chemistry more balanced vs + positives
Feature No specific ; more dedicated to difficult cells

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Cell Culture surfaces - Adherent Cells

Synthetic Non Specific CellBIND & Primaria
CellBIND

TCT

BD
Primaria

Treatment

N2O

O2

NH4 /proprietary

% Oxygen/
Nitrogen

35% Oxygen

22% Oxygen

14.1% Oxygen
11.1% Nitrogen

1.8% Oxygen

Contact
angle

23 - 26

56 - 63

75

90 - 95

Charge

Negative

Negative

Positive

Not Treated

TCT

CellBIND
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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
Biological coating with ECM molecules :
Cells are not living in 2D into a plastic box !
They are normally living in the context of other cells, surrounded and in
contact with an extracellular matrix (ECM)
The TC-treated surfaces can be coated with ECM components :
Biological molecules from the ECM :
Can be bought pre-coated ready-to-use products.
Can be bought as a chemical to plate into the lab.
Reconstituted ECM = Corning Matrigel Matrix :
Can be bought pre-coated ready-to-use products.
Can be bought as a chemical to plate into the lab.

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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
READY-TO-USE
Biological molecules from the ECM :

Collagen I
Collagen IV
Fibronectin
Gelatin
Laminin
Poly-Lysine
Poly-D-Lysine/Laminin
Poly-L-Ornithine/Laminin
Laminin/Fibronectin

Reconstituted ECM = Corning Matrigel Matrix :

Matrigel
Matrigel Growth Factors Reduced (GFR)
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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
TO COAT INTO THE LAB :
Biological molecules from the ECM :

Collagen I, II, III, IV, V, VI

Fibronectin
Laminin
Osteopontin
Poly-Lysine
Vitronectin

Reconstituted ECM = Corning Matrigel Matrix :

Matrigel
Matrigel hESC (human Embryonic Stem Cells) qualified
Matrigel HC to do tumor injection
Matrigel Phenol Red free
Matrigel GFR (Growth Factor Reduced)
Dispase & Cell Recovery Solution to recover cells from Matrigel
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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
APPLICATIONS :

Endothelial cell migration, invasion

Angiogenesis tube formation
Tumor invasion
3-D-cell culture
Maintenance of human embryonic stem cells
Neurite outgrowth
Hepatocyte culture

SEMs of hepatocytes cultured for two days on collagen I (1),

collagen I gels (2), or BD Matrigel Matrix (3). Note the cluster of
spherical cells for hepatocytes cultured on Corning Matrigel Matrix,
typical of differentiated cells.
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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
APPLICATIONS :
HEK293

Before Wash

After Wash

After Wash,
Calcein AM Staining

After Wash

After Wash,
Calcein AM Staining

TC Plates

Before Wash

BD BioCoat
PDL Plates

HEK-293 cells were cultured under serum-free conditions on TC-treated or BD

BioCoat PDL 384-well black/clear plates. Samples were washed using a
Skatron Washer (Molecular Devices).
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Cell Culture surfaces - Adherent Cells

ECM Biological Coating
ADVANTAGES :
More in vivo like properties
Better way of life :
attachment, proliferation, growth
including difficult cells as : primary cells, neural cells, hepatocytes,
endothelial cells,
Differentiation and functional studies possible
3-dimensional culture of cells possible, when cells are embedded in a thick gel,
and not only applied to a thin coating
Testing packs to try different molecules (mix of different surfaces)

LIMITATION :
Not Animal components free
Limit shelf life
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Cell Culture surfaces - Adherent Cells

Synthetic defined - Purecoat
An advanced cell culture surfaces, that uses a novel process to
coat the culture vessel in a very defined and reproducible way
2 options :
amine (+) residues
carboxyl (-) residues
BD PureCoatTM

Advantages :

Chemically defined
Animal-free
High consistency
Low CVs in cell-based assays
Stable at room temperature (3 years)

+ + + + + +
or
- - - - - - -

Thin-film coating
Carboxyl or Amine
momomers

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Cell Culture surfaces - Adherent Cells

Synthetic defined - Purecoat

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Cell Culture surfaces - Adherent Cells

Synthetic defined Purecoat ECM Mimetic
2 options :
Fibronectine
Collagen

ECM Mimetic Surface technology

Main applications :
Human Mesenchymal Stem Cells
Human Endothelial Progenitor Cells
Human Keratinocytes

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Cell Culture surfaces - Adherent Cells

Synthetic defined PuraMatrix
PuraMatrix Petpide Hydrogel :
-

Synthetic matrix to create a defined 3D microenvironments

origin: synthetic, animal free
major components: - 16 aa peptide
average pore size 50-200 nm
no growth factors

- Applications :
-

Puramatrix scaffold allows creation of highly defined environment through

addition of factors & compounds
Does not activate RGD-dependent integrin signaling
Highly reproducible experiments
Compatible with most cells types in vitro & in vivo (stem cell proliferation, tumor
cell migration and invasion, angiogenesis assays, tissue regeneration)

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Cell Culture surfaces - Adherent Cells

Synthetic Specific Synthemax
Synthemax Surface :
1 Application Stem cell
What is the Corning Synthemax Surface ?

polystyrene

1. Uncoated surface

polystyrene

2. Acrylate coated

polystyrene

3. Peptide covalently
bound

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Cell Culture surfaces - Adherent Cells

Synthetic Specific Synthemax
Advantages : Time Saving & Shelf Life
Feeders

Storage

Shelf Life

Easy-to-Use

ECMs

Synthemax

Need to be used
within a week

Need to be stored at 70C

When working they
need to be kept cold

No special storage
conditions; can be
stored at room temp

Last only a week once

prepared

Plates need to be used

as soon as possible

2 year shelf life

Harvest mouse
embryos
Grow up the
Fibroblasts
Time intensive

Laborious timeconsuming prep

No prep, just cut

open the package and
use

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Cell Culture surfaces - Adherent Cells

Synthetic Specific Synthemax
Customer Needs

Research

Therapeutic

Traceability

10-6 Sterility

Consistent Performance

Screening

Shelf Life
No Special Storage Conditions

Ease of Use

Purity Post-Harvesting

Lot Size

Multiple Formats

Inventory Availability

Xeno-Free Substrate

Ability to visualize cells

Automation Friendly Format

GMP Production
Expand & Differentiate on same Surface

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Cell Culture surfaces - Adherent Cells

Synthetic Specific Synthemax

Synthemax

Consistency

Defined, Synthetic,
Xeno-Free

Manufactured under GMP conditions

Performs as good as Matrigel
Consistent performance
2 year shelf life
Defined
Synthetic
No animal components
No prep, just cut open the package and use

Easy-to-Use

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Cell Culture surfaces - Adherent Cells

Synthetic Specific Osteo Assay
Osteo Assay Surface :
1 Application Osteo/bone assays

Specifications :
Inorganic synthetic coating that mimics
in-vivo bone for in vitro bone cell assays
Surface applications :
- Consistent assay results for bone cell
- Differentiation and function
- Co-culture of bone cells / metastatic cancer cells
- Alternative to preparing dentine, bone slices
SEM image
magnified not true
representation
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Cell Culture surfaces - Adherent Cells

Synthetic Specific Osteo Assay
Key Benefits

Critical Attributes

Differentiating Attributes

Time

Reduce time and cost of surface

preparation

Precoated, sterile, ready to use

Direct Readout

Detect both cell differentiation and

functional analysis of bone cells on the
same surface
Supernatant can be sampled for cell
function

Transparent to conventional imaging

techniques

Amenable to
automation

Standard SBS format

Can be used with standard imaging

technologies

Reliable cell
response

Consistent assays

Standard process for manufacturing

Uniform surface

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Cell Culture surfaces Procuts available

You can find the different cell culture surfaces on multiple vessels :
Flasks
Microplates from 6 to 1536 wells
Dishes

For specific applications :

Assays microplates :
Drug discovery
Molecules screening

Cells Culture inserts (Transwell) :

Coculture
Migration
Invasion

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TOOLS TO FIND THE RIGHT SURFACE

Cell Culture Selection Guide Corning

Cell Culture Selection Guide Corning Falcon
Success Stories CellBIND
Bibliography OsteoAssays
Bibliography ULA (Ultra Low Attachment)

Different Protocols or Application notes

European Scientific Support (2 scientists)

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Glass Bottom Microplates

Glass
High optical quality and
scratch-resistant glass
Bottom thickness 200 m
Intra well bottom flatness <50
m

Copolimer
Cyclic Olefin Copolymer
(COC)
Bottom thickness: 127 m
Intra-well bottom flatness 5
m
Inter-well bottom flatness 30
m

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Flowell

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Flowell

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Axygen 5 mL Snaplock Microtubes

5mL
15mm

No necesita adaptador
Rotores 15 mm (15ml)
Rotores 17 mm (15ml)

14.000 rpm
120C (15 15psi)
Libre de ARNasas, ADNasas

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Corning Life Science - AXYGEN Biosciences

Thanks a lot for your time and attention.

QUESTIONS ?

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