Transwell®

Permeable Supports
Including Snapwell™ and
Netwell™ Inserts

Corning Incorporated Worldwide EUROPE

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Instructions for Use
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Corning and Transwell are registered trademarks of Corning, Incorporated, Corning, NY.
Netwell and Snapwell are trademarks of Corning, Incorporated, Corning, NY.
Corning Incorporated, One Riverfront Plaza, Corning, NY 14831-0001
TA B L E O F C O N T E N T S INTRODUCTION
Introduction ....................................1 Transwell® Permeable Supports are convenient and easy-to-use
devices for studies of both anchorage-dependent and anchorage-
Transwell® Insert Product Descriptions . . . . . . . . . . . . . 1 independent cells. These inserts provide independent access to
Standard Transwell Inserts . . . . . . . . . . . . . . . . . . . . . 1 both sides of a monolayer, thus giving researchers a versatile tool
to study transport and other metabolic activities in vitro. Transwell
Membrane Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 units are available in a range of diameters, membrane types and
Pore Sizes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 pore sizes to satisfy a variety of research requirements.
Pore Density . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Chemical Compatibility . . . . . . . . . . . . . . . . . . . . . . . 4 T R A N SW E L L I N S E RT P R O D U C T D E S C R I P T I O N S
Using Transwell Permeable Supports . . . . . . . . . . . . . . . 5
Directions for Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Helpful Hints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

Additional Permeable Support Products . . . . . . . . . . . . . 7

HTS Transwell-24 Well Permeable Supports . . . . . . 7
HTS Transwell-96 Well Permeable Supports . . . . . . 8 Standard Transwell Inserts
Snapwell™ Inserts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 Standard Transwell inserts are available in four membrane
Netwell™ Inserts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 diameters: 6.5 mm (24 well plate), 12 mm (12 well plate), 24 mm
(6 well plate) and 75 mm (100 mm dish) formats. See Table 1 for
Technical Assistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 cell growth areas.

Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Table 1. Transwell Permeable Support Growth Areas*

Polycarbonate Transwell Inserts . . . . . . . . . . . . . . . . 10 Transwell Multiple Well Plate Insert Membrane
Insert Diameter or Dish Style Growth Area
Polyester Transwell Inserts . . . . . . . . . . . . . . . . . . . . 10
6.5 mm 24 well 0.33 cm2
Collagen Coated Transwell-COL Inserts . . . . . . . . . 11 12 mm 12 well 1.12 cm2
HTS Transwell 24 System . . . . . . . . . . . . . . . . . . . . 11 24 mm 6 well 4.67 cm2
HTS Transwell-96 System . . . . . . . . . . . . . . . . . . . . 12 75 mm 100 mm dish 44 cm2
Snapwell Inserts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 * Values are reported as nominal and may vary slightly due to inherent variability
of our manufacturing process. To insure success, we recommend that end users
Netwell Inserts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 validate their methods independent from our reported values.
6, 12, and 24 Well Cell Culture Plates . . . . . . . . . . . 13

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Several membrane Polycarbonate Transwell® inserts feature thin, translucent mem-
types and a large Transwell® insert branes available in six pore sizes ranging from 0.1 µm to 12.0 µm.
selection of pore Most are treated for optimal cell attachment. They are supplied
sizes are available Upper compartment sterile and come preloaded in multiple well plates or dishes.
with each of these Microporous membrane The polycarbonate membrane is compatible with most organic
units. A patented* Lower compartment fixatives and stains.
self-centering Polyester (PET) Transwell-Clear inserts have microscopically
design prevents transparent polyester membranes that are tissue culture treated
medium from for optimal cell attachment and growth. Transwell-Clear inserts
wicking between the sides of the insert and the well wall. The provide better cell visibility under phase contrast microscopy and
hanging design keeps the Transwell® membrane about a milli- allow assessment of cell viability and monolayer formation.
meter off the bottom of the well. This prevents co-cultured cell
monolayers in the bottom of the well from being scratched or Transwell-COL inserts have transparent (when wet), collagen-
disturbed when the insert is moved. Windows or openings in the treated PTFE membranes that promote cell attachment and
sides of the Transwell insert allow access to the lower compartment. spreading and allow cells to be visualized during culture. The
Transwell-COL membranes are coated with an equimolar mix-
Membrane Types ture of types I and III collagen derived from bovine placentas.
Transwell permeable supports are available in three membrane Unlike traditional coating techniques that result in occluding
materials: polycarbonate, polyester (PET), and collagen-coated film layers, Corning’s proprietary coating process results in a
polytetrafluoroethylene (PTFE). See Table 2 for additional biologically stable collagen that covers every fibril of the filter
information on these membrane characteristics. matrix, thereby retaining the porosity of the membrane.

Pore Sizes
Table 2. Characteristics of Transwell Membranes
Polyester PTFE/
Selecting the correct pore size for experiments is also very impor-
Characteristics (PET) Polycarbonate Collagen-coated tant. The smaller pore size Transwell membranes (0.4 or 3.0 µm)
Optical properties Clear Translucent Clear when wet are primarily used in drug transport studies. Cell invasion, chemo-
Cell visibility Good Poor Cell outlines taxis and motility studies are usually done in Transwell membranes
Tissue culture treated Yes Yes No
with 3.0 µm or larger pores. The ability of cells to migrate
through pores of a membrane is dependent on the cell line used
Nominal Membrane 10 µm 10 µm 30 µm
thickness‡
and the culture conditions, as well as the pore size. Cell migra-
tion will not occur with pores smaller than 3.0 µm. For critical
Matrix/ECM coatable Yes Yes Yes
experiments, we suggest testing with appropriate controls and a
Collagen treated No No Yes
range of pores sizes to determine which size works best with your
Available Pore Sizes (µm) 0.4, 1.0, 0.4, 3.0, 0.4, 3.0 cell cultures and your specific application. As an alternative, fol-
3.0, 8.0 5.0, 8.0
low recommendations from the scientific literature. For additional
Membrane manufacturing processes are inherently variable; thus, actual values may vary up to 60% of nominal
‡

depending on membrane type and measurement techniques. application and use information, please refer to the Transwell
*Patent number 5,026,649.

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inserts bibliography on the Technical Information section of the U S I N G T R A N S W E L L® P E R M E A B L E S U P P O R T S
Corning Life Sciences web site (www.corning.com/ lifesciences)
that lists over 800 literature references using Transwell permeable Directions for Use
supports. 1. Transwell inserts are used by first adding medium to the multiple
well plate well, followed by adding the Transwell inserts, and
Pore Density lastly adding the medium and cells to the inside compartment.
Of the three types of Transwell® membranes, only the PTFE Recommended medium volumes are shown in Table 4.
membrane does not have a defined pore density because it is a
tortuous path membrane. The two membranes with nominally Table 4. Recommended Transwell Permeable Support
defined pore densities are polycarbonate and polyester. The poly- Medium Volumes
ester Transwell membranes do not have as high a pore density as Transwell Insert Multiple Well Volume Volume Added
Insert Membrane Plate or Added per to Inside of
the polycarbonate Transwell membranes but they have better Diameter Growth Area Dish Type Plate Well Transwell Insert
optical clarity as a result. The nominal pore densities for poly- 4.26 mm 0.143 cm2 96 well 0.235 mL 0.075 mL
carbonate and polyester membranes are given in Table 3. 6.5 mm 0.33 cm2 24 well 0.6 mL 0.1 mL
12 mm 1.12 cm2 12 well 1.5 mL 0.5 mL
Table 3. Nominal Pore Densities for Transwell Polyester
24 mm 4.67 cm2 6 well 2.6 mL 1.5 mL
and Polycarbonate Membranes
75 mm 44 cm2 100 mm dish 13 mL 9.0 mL
Nominal Pore Density
Pore Size Polycarbonate Membrane Polyester Membrane 2. An initial equilibrium period may be used to improve cell
0.4 µm 1 x 108 pores/cm2 4 x 106 pores/cm2 attachment by adding medium to the multiple well plate well
1.0 µm – 1.6 x 106 pores/cm2 and then to the Transwell insert. The plate should then be
3.0 µm 2 x 106 pores/cm2 2 x 106 pores/cm2 incubated for at least one hour or even overnight at the same
5.0 µm 4 x 105 pores/cm2 –
temperature that will be used to grow the cells. The cells are
then added in fresh medium to the Transwell insert and
8.0 µm 1 x 105 pores/cm2 1 x 105 pores/cm2
returned to the incubator.
Chemical Compatibility 3. The medium level should be checked periodically and fresh
All of the Transwell membranes are compatible with histological medium added as required.
fixatives including methanol and formaldehyde. The polyester 4. Transwell inserts have three openings for standard pipette tips
Transwell membranes have the best overall chemical resistance. to allow samples to be added or removed from the lower
These membranes (but not the polystyrene housings) are com- compartment.
patible with many alcohols, amines, esters, ethers, ketones, oils 5. Cell monolayers may be fixed and stained while in the Transwell
and some solvents including many halogenated hydrocarbons and insert using standard cytological techniques. Avoid using sol-
DMSO, but are not recommended for use with strong acids and vents that dissolve polystyrene or polycarbonate or polyester
bases. membrane materials. Processing steps may be carried out by

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 sequentially moving the Transwell insert through a series of 5. Cells requiring extracellular matrix coatings on plastic
multiple well plate wells containing the appropriate reagents. substrates will also require them on permeable supports.
Protocols for fixing and staining Transwell® inserts are 6. The Transwell®-Clear insert contains a transparent tissue
available on the Corning Life Sciences web site culture treated polyester membrane that allows easy viewing
(www.corning.com/lifesciences). of cells using phase contrast microscopy.
6. If it is necessary to remove cells from Transwell membranes, 7. The Transwell-COL insert contains a PTFE membrane that
we recommend rinsing both the Transwell insert and the plate has been treated with an equimolar mixture of types I and III
well. Then the dissociating solution should be added to both bovine placental collagens. This results in a biologically
the well and the Transwell insert and incubated until the cells stabilized collagen matrix covering the fibrils of the filter
begin to come off. A protocol, Trypsinization Procedure for membrane. These Transwell inserts are excellent for the
Corning® Transwell® Inserts, for removing cells from Transwell growth of cells requiring a biological coating.
inserts is available in the technical section of the Corning Life
Sciences web site.
ADDITIONAL PERMEAB LE SU PPORT
7. The polycarbonate or polyester membrane with the fixed and
stained cells attached may be removed from the Transwell insert PRODUCTS
by carefully cutting around the membrane edges with a scalpel.
HTS Transwell-24 System
8. The collagen-coated PTFE membrane is fragile and requires
careful handling during removal. A wetted cellulosic membrane The HTS Transwell-24
filter should be placed in direct contact with the underside System is an array of 24 indi-
of the Transwell insert membrane before it is cut out with a vidual Transwell inserts (6.5
scalpel. The wetted, more rigid, cellulosic filter will serve as mm diameter membranes)
a support for the collagen-coated membrane. connected by a rigid, robotics-
friendly holder that enables
Helpful Hints all 24 Transwell inserts to be
1. Cell morphology and cell densities on permeable supports handled as a single unit. This
are influenced by filter pore size. makes the HTS Transwell-24
System an ideal tool for running automated, high throughput
2. Larger pore sizes may permit some cell types to migrate drug transport (Caco-2 cells) or cell toxicity studies. The tissue
through the pores on the permeable support. culture treated polycarbonate membrane is available in either
3. Cells grown on permeable supports are often sensitive to 0.4 µm or 3.0 µm pore sizes. The tissue culture treated polyester
initial seeding density for good cell attachment. On first use, try membrane is available in 0.4 µm pore size. Both membrane types
bracketing a range of seeding densities for optimum growth. provide an excellent substrate for cell attachment, growth, and
4. Cell attachment and spreading may be improved by differentiation. An open culture reservoir plate is used to reduce
preincubating permeable supports in medium prior to seeding. liquid handling during cell feeding (medium can be exchanged all
at once). Once the cell layers are confluent, the HTS Transwell-24
insert is transferred to a 24 well plate for running experiments.

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HTS Transwell®-96 Permeable Support Systems Netwell™ Inserts
and Plates Netwell inserts are polystyrene inserts fitted with polyester
The HTS Transwell-96 product is an array of 96 Transwell mesh bottoms for use with 6 and 12 well plates.
inserts integrated into a single robotics-friendly unit. Plates are Typical Applications
available with either polyester (PET) membrane or polycarbon-
◗ For 3-D tissue culture, the sterile Netwell insert provides
ate (PC) membrane.
efficient media permeation and support to cultures grown
All standard units are tissue culture treated (membrane only) and in gels
sterile. The HTS Transwell-96 Permeable Supports have a well
◗ Sterile Netwell inserts provide easy processing of tissue
bottom elevation of 1.3 mm.
explants and microcarrier-grown cells
The HTS Transwell-96 Systems (0.4 µm PC and 1.0 µm PET) ◗ Modular design makes the Netwell insert handy for
are designed for drug transport, excretion, and other ADMET immunocytochemical staining on multiple sections
assays. The systems are packaged with the 96 well insert plate in
a reservoir plate and includes the 96 well receiver plate with lid.
The HTS Transwell-96 Well Plates (3.0 µm PC, 5.0 µm PC, T E C H N I C A L A S S I S TA N C E
8.0 µm PET) are ideal for chemotaxis, migration, and invasion For additional product or technical information, please visit
assays. These are packaged with the 96 well insert plate in the www.corning.com/lifesciences or call 1.800.492.1110. The
96 well receiver plate with lid. web site contains protocols and a bibliography with over 800
Snapwell™ Inserts references for using Transwell® permeable supports. Customers
outside the United States, please call +1.978.635.2200.
The Snapwell insert is a modified
Transwell culture insert that con-
tains a 12 mm diameter tissue
culture treated polycarbonate or
clear polyester membrane sup-
ported by a detachable ring. These
inserts are primarily used for
transport and electrophysiological
studies. Once cells are grown to
confluence, this ring-supported
membrane can be placed into either vertical or horizontal
diffusion or Ussing chambers. Chambers are available from
Harvard Apparatus, www.harvardapparatus.com.
*Patent numbers 5,026,649 and 5,139,951.

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O R D E R I N G I N F O R M AT I O N

Polycarbonate Membrane Transwell® Inserts Collagen-Coated Transwell®-COL Inserts

Membrane Growth Membrane Tissue Membrane Growth Membrane
Diameter Surface Pore Culture Inner Inserts/ Cat. Diameter Surface Pore Size Inner Multiple Inserts/
Cat. No. (mm) Area (cm2) Size (µm) Treated Packaging Case No. (mm) Area (cm2) (µm) Packaging Well Plate Case
3413 6.5 0.33 0.4 Yes 12/plate* 48 3495 6.5 0.33 0.4 Individual 24 well 24
3415 6.5 0.33 3.0 Yes 12/plate* 48 3496 6.5 0.33 3.0 Individual 24 well 24
3421 6.5 0.33 5.0 Yes 12/plate* 48 3493 12 1.12 0.4 Individual 12 well 24
3422 6.5 0.33 8.0 Yes 12/plate* 48 3494 12 1.12 3.0 Individual 12 well 24
3401 12 1.12 0.4 Yes 12/plate 48 3491 24 4.67 0.4 Individual 6 well 24
3402 12 1.12 3.0 Yes 12/plate 48 3492 24 4.67 3.0 Individual 6 well 24
3412 24 4.67 0.4 Yes 6/plate 24
3414 24 4.67 3.0 Yes 6/plate 24 HTS Transwell-24 Systems
3428 24 4.67 8.0 Yes 6/plate 24 Cat. Pore Size Qty/ Qty/
3419 75 44 0.4 Yes 1/dish 12 No. Description (µm) Pk Cs
3420 75 44 3.0 Yes 1/dish 12 3378 HTS Transwell-24, PET membrane, Bulk 0.4 12 12
*Packaged 12 inserts per 24 well plate 3379 HTS Transwell-24, PET membrane, 0.4 1 2
Individual
3395 HTS Transwell Open Culture Reservoir N/A 12 48
Polyester (PET) Membrane Transwell-Clear Inserts
3396 HTS Transwell-24, Individual 0.4 1 2
Membrane Growth Membrane
Cat. Diameter Surface Pore Size Inner Inserts/ 3397 HTS Transwell-24, Bulk 0.4 12 12
No. (mm) Area (cm2) (µm) Packaging Case 3398 HTS Transwell-24, Individual 3.0 1 2
3470 6.5 0.33 0.4 12/plate* 48 3399 HTS Transwell-24, Bulk 3.0 12 12
3472 6.5 0.33 3.0 12/plate* 48
3460 12 1.12 0.4 12/plate 48
3462 12 1.12 3.0 12/plate 48
3450 24 4.67 0.4 6/plate 24
3452 24 4.67 3.0 6/plate 24
*Packaged 12 inserts per 24 well plate

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O R D E R I N G I N F O R M AT I O N (CONTINUED)

HTS Transwell®-96 Well Permeable Supports Netwell™ Inserts

Pore Membrane Membrane
Cat. Size Qty/ Qty/ diameter Mesh Size Inner Inserts/
No. Description (µm) Membrane Pk Cs Cat. No. (mm) (µm) Sterile Packaging Case
3380 HTS Transwell-96 System 1.0 PET 1 1 3477 15 74 Yes 12/plate 48
3392 HTS Transwell-96 System 1.0 PET 1 5 3478 15 500 Yes 12plate 48
3381 HTS Transwell-96 System 0.4 PC 1 1 3479 24 74 Yes 6/plate 48
3391 HTS Transwell-96 System 0.4 PC 1 5 3480 24 500 Yes 6/plate 48
3385 HTS Transwell-96 Well Plate 3.0 PC 1 2
3386 HTS Transwell-96 Well Plate 3.0 PC 4 8 Netwell™ Accessories
3387 HTS Transwell-96 Well Plate 5.0 PC 4 8 Cat. No. Description Inserts/Case
3388 HTS Transwell-96 Well Plate 5.0 PC 1 2 3517 Netwell reagent tray, black 200
3374 HTS Transwell-96 Well Plate 8.0 PET 1 2 3519 Netwell reagent tray, white 200
3384 HTS Transwell-96 Well Plate 8.0 PET 4 8 3520 Netwell carrier kit*, 15 mm 8
3382 HTS Transwell-96 Receiver N/A N/A 10 10 3521 Netwell carrier kit*, 24 mm 8
Plate with lid
*Kits contain eight carriers and eight handles.
3383 HTS Transwell-96 Feeder N/A N/A 10 10
Reservoir with media stabilizer
and lid 6, 12, and 24 Well Cell Culture Plates
3583 HTS Transwell-96 black N/A N/A 10 10 Number Well Growth
receiver plate with lid, TCT Cat. No. of Wells Diameter (mm) Area (cm2) Qty/Pk Qty/Cs
3506 6 34.8 9.5 5 100
Snapwell Inserts
™ 3516 6 34.8 9.5 1 50
3512 12 22.1 3.8 5 100
Membrane
Pore Size Membrane Inner Inserts/ 3513 12 22.1 3.8 1 50
Cat. No. (µm) Material Packaging Case
3524 24 15.6 1.9 1 100
3407 0.4 Polycarbonate 6/plate 24
3526 24 15.6 1.9 1 50
3801 0.4 Clear Polyester 6/plate 24
3527 24 15.6 1.9 5 100
3802 3.0 Polycarbonate 6/plate 24

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