Epithelial Cells - PromoCell

Epithelial Cells

Instruction Manual Size Catalog Number

Product 500,000 cryopreserved cells C-12620
500,000 proliferating cells C-12621
Human Nasal Epithelial Cells
(HNEpC) 500,000 cryopreserved cells C-12644
Human Tracheal Epithelial Cells 500,000 proliferating cells C-12645
(HTEpC)
Human Bronchial Epithelial Cells 500,000 cryopreserved cells C-12640
(HBEpC) 500,000 proliferating cells C-12641
Human Small Airway Epithelial Cells
(HSAEpC) 500,000 cryopreserved cells C-12642
Human Renal Epithelial Cells 500,000 proliferating cells C-12643
(HREpC)
Human Renal Cortical Epithelial Cells 500,000 cryopreserved cells C-12665
(HRCEpC) 500,000 proliferating cells C-12666
Human Placental Epithelial Cells
(HPlEpC) 500,000 cryopreserved cells C-12660
500,000 proliferating cells C-12662

500,000 cryopreserved cells C-12395
500,000 proliferating cells C-12397

Product Description 500,000 cryopreserved cells that have Intended Use
been thawed and cultured for three days
The epithelium is the interface between at PromoCell. PromoCell Epithelial Cells are for in vitro
the body and the external environment research use only and not for diagnostic
and covers all exterior surfaces and in- Quality Control or therapeutic procedures.
terior lumina. Depending on the tissue
of origin, the functions of epithelial cells Rigid quality control tests are performed Warning
are diverse and include absorption, se- for each lot of Epithelial Cells.
cretion, protection, transcellular trans- They are tested for cell morphology, Although tested negative for HIV-1,
port, and sensation. adherence rate and cell viability. Fur- HIV-2, HBV, HCV, HTLV-1 and HTLV-2,
PromoCell offers a range of Epithelial thermore, flow cytometric analyses for the cells – like all products of human
Cells produced at PromoCell’s cell cul- the cell-type specific marker cytokeratin origin – should be handled as potentially
ture facility. The cells are isolated from are carried out for each lot (see page 5). infectious. No test procedure can com-
normal human adult tissues of the air- Growth performance is tested through pletely guarantee the absence of infec-
ways (nasal mucosa, trachea, bronchi, multiple passages under culture condi- tious agents.
and distal respiratory tract), kidney tions without antibiotics and antimycotics.
(whole kidney and renal cortex), and the In addition, all cells have been tested for Follow appropriate safety precautions!
placenta (amniotic membrane). the absence of HIV-1, HIV-2, HBV, HCV,
Shortly after isolation, all PromoCell HTLV-1, HTLV-2 and microbial contami- After delivery, start immediately with
Human Epithelial Cells are cryopreserved nants (fungi, bacteria, and mycoplasma). the protocol for cryopreserved cells (see
at passage 2 (P2) using PromoCell´s pro- A detailed certificate of analysis (CoA) page 2) or the protocol for proliferating
prietary, serum-free freezing medium, for each lot can be downloaded at: cells (see page 3).
Cryo-SFM. Each cryo vial contains more www.promocell.com/coa
than 500,000 viable cells after thawing.
Proliferating cell cultures are made from

2 Instruction Manual - Epithelial Cells

Start immediately after delivery.
Use aseptic techniques and a laminar flow bench.

Protocol for Cryopreserved Cells

Straight after arrival, store the cryopreserved cells in liquid nitrogen or seed them immediately.
Note: Storage at -80°C is not sufficient for cell preservation and causes irreversible cell damage.

1. Prepare the medium
Calculate the needed culture surface
area according to the plating density
(see page 5) and the lot-specific cell
numbers stated on the certificate of
analysis. Fill the appropriate volume
of PromoCell Growth Medium (at
least 9 ml per vial of cells) in cell
culture vessels. Place the vessels in
an incubator (37°C, 5% CO2) for 30
minutes.

2. Thaw the cells
Remove the cryovial from the liquid
nitrogen container and immediately
place it on dry ice – even for short
transportation. Under a laminar flow
bench, briefly twist the cap a quar-
ter turn to relieve pressure, then re-
tighten. Immerse the vial into a water
bath (37°C) just up to the screw cap
for 2 minutes. Ensure that no water
enters the thread of the screw cap.

3. Disinfect the vial and seed the cells
Thoroughly rinse the cryovial with
70% ethanol under a laminar flow
bench. Then, aspirate the excess
ethanol from the thread area of the
screw cap. Open the vial and transfer
the cells to a cell culture vessel con-
taining the prewarmed medium from
step 1.

4. Incubate the cells
Place the vessel in an incubator (37°C,
5% CO2) for cell attachment. Replace
the medium after 16-24 hours. The
cells should be subcultured according
to the subcultivation protocol (see
page 4) once they have reached
70 – 90% confluency.

Instruction Manual - Epithelial Cells 3

Start immediately after delivery.
Use aseptic techniques and a laminar flow bench.

Protocol for Proliferating Cells

1. Incubate the cells
Unpack the culture vessel, do not
open the lid, and immediately place
it in an incubator (37°C, 5% CO2) for
3 hours to allow the cells to recover
from the transportation.

2. Replace the transport medium
Carefully open the vessel, rinse
the inner side of the lid with 70%
ethanol, and let air dry. Aspirate the
transport medium from the vessel.
Add 10 ml of the appropriate Promo-
Cell Cell Growth Medium.

3. Check and incubate the cells
Check the cell density. Open the lid
half a turn and place the vessel in an
incubator (37°C, 5% CO2). The cells
should be subcultured according to
the subcultivation protocol (see page
4) once they have reached 70 – 90%
confluency.

Instruction Manual - Epithelial Cells 4

Use aseptic techniques and a laminar flow bench.

Subcultivation Protocol

1. Prepare the reagents and wash
the cells
Place the PromoCell DetachKit at room
temperature for at least 30 minutes to
adjust the temperature of the reagents.
Carefully aspirate the medium from
the culture vessel. Add 100 µl Hepes
BSS Solution per cm2 of vessel surface
to wash the cells and agitate the
vessel carefully for 15 seconds.

2. Detach the cells
Carefully aspirate the Hepes BSS from
the culture vessel. Add 100 µl Trypsin/
EDTA Solution per cm2 of vessel sur-
face. Note: We recommend detach-
ing the cells at room temperature. Close
the vessel and examine the cells under
a microscope. When the cells start to
detach, gently tap the side of the ves-
sel to loosen the remaining cells.

3. Neutralize the trypsin and harvest
the cells
Add 100 µl Trypsin Neutralization Solu-
tion per cm2 of vessel surface and
gently agitate. Carefully aspirate the
cell suspension and transfer it to a
centrifugation tube. Spin down the
cells for 3 minutes at 220 x g.

4. Incubate the cells
Discard the supernatant (step 1), add
1 ml of the appropriate PromoCell Cell
Growth Medium (step 2), and resus-
pend the cells by carefully pipetting up
and down. Plate the cells according
to the recommended seeding density
in new cell culture vessels containing
PromoCell Cell Growth Medium pre-
warmed to 37°C. Place the vessels in
an incubator (37°C, 5% CO2).

5 Instruction Manual - Epithelial Cells

Specifications

Product Recommended Plating Density Passage after Marker Population
Culture Media* Thawing Doublings
Human Nasal
Epithelial Cells (HNEpC) C-21060 10,000 - 15,000 P2 Cytokeratin+ > 15
cells per cm² P2 Cytokeratin+
Human Tracheal C-21060 P2 Cytokeratin+ > 15
Epithelial Cells (HTEpC) 10,000 - 15,000 P2 Cytokeratin+
C-21060 cells per cm² P2 Cytokeratin+ > 15
Human Bronchial P2 Cytokeratin+
Epithelial Cells (HBEpC) C-21070 10,000 - 15,000 P2 Cytokeratin+ > 15
cells per cm²
Human Small Airway C-26001 > 15
Epithelial Cells (HSAEpC) C-26030 10,000 - 15,000
C-26001 cells per cm² > 15
Human Renal C-26030
Epithelial Cells (HREpC) C-26040 10,000 - 15,000 >5
cells per cm²
Human Renal Cortical Catalog Number
Epithelial Cells (HRCEpC) 10,000 - 15,000
cells per cm² C-21060
Human Placental C-21160
Epithelial Cells (HPlEpC) 10,000 - 15,000 C-21260
cells per cm² C-21265
C-39165
Related Products Size C-39160
C-21070
Product 500 ml C-21170
500 ml C-21270
Airway Epithelial Cell Growth Medium (Ready-to-use) 500 ml C-21275
Airway Epithelial Cell Growth Medium Kit 500 ml C-39175
Airway Epithelial Cell Basal Medium for 500 ml C-39170
Airway Epithelial Cell Basal Medium, phenol red-free for 500 ml C-21010
Airway Epithelial Cell Growth Medium SupplementMix C-21110
Airway Epithelial Cell Growth Medium SupplementPack 500 ml C-21210
500 ml C-21215
Small Airway Epithelial Cell Growth Medium (Ready-to-use) 500 ml C-39115
Small Airway Epithelial Cell Growth Medium Kit 500 ml C-39110
Small Airway Epithelial Cell Basal Medium for 500 ml
Small Airway Epithelial Cell Basal Medium, phenol red-free for 500 ml
Small Airway Epithelial Cell Growth Medium SupplementMix 500 ml
Small Airway Epithelial Cell Growth Medium SupplementPack 500 ml
Mammary Epithelial Cell Growth Medium (Ready-to-use) 500 ml
Mammary Epithelial Cell Growth Medium Kit 500 ml
Mammary Epithelial Cell Basal Medium for 500 ml
Mammary Epithelial Cell Basal Medium, phenol red-free for 500 ml
Mammary Epithelial Cell Growth Medium SupplementMix
Mammary Epithelial Cell Growth Medium SupplementPack

*The catalog numbers in this table are for media in ready-to-use packaging.

Product Size Catalog Number

Renal Epithelial Cell Growth Medium (Ready-to-use) 500 ml C-26001
Renal Epithelial Cell Growth Medium Kit 500 ml C-26011
Renal Epithelial Cell Basal Medium 500 ml C-26021
Renal Epithelial Cell Basal Medium, phenol red-free 500 ml C-26022
Renal Epithelial Cell Growth Medium SupplementMix for 500 ml C-39604
Renal Epithelial Cell Growth Medium SupplementPack for 500 ml C-39600
Renal Epithelial Cell Growth Medium 2 (Ready-to-use) 500 ml C-26030
Renal Epithelial Cell Growth Medium 2 Kit 500 ml C-26130
Renal Epithelial Cell Basal Medium 2 500 ml C-26230
Renal Epithelial Cell Basal Medium 2, phenol red-free 500 ml C-26235
Renal Epithelial Cell Growth Medium 2 SupplementMix for 500 ml C-39606
Renal Epithelial Cell Growth Medium 2 SupplementPack for 500 ml C-39605

Placental Epithelial Cell Growth Medium (Ready-to-use) 500 ml C-26040
Placental Epithelial Cell Growth Medium Kit 500 ml C-26140
Placental Epithelial Cell Basal Medium 500 ml C-26240
Placental Epithelial Cell Basal Medium, phenol red-free
Placental Epithelial Cell Growth Medium SupplementMix 500 ml C-26245
Placental Epithelial Cell Growth Medium SupplementPack for 500 ml C-39612
for 500 ml C-39610
DetachKit
30 ml C-41200
Cryo-SFM 125 ml C-41210
250 ml C-41220
HNEpC Pellet C-29910
HTEpC Pellet 30 ml C-29912
HBEpC Pellet 125 ml C-14062
HSAEpC Pellet 1 million cells per pellet C-14064
HREpC Pellet 1 million cells per pellet C-14063
HRCEpC Pellet 1 million cells per pellet C-14065
HPlEpC Pellet 1 million cells per pellet C-14067
BPE-15 1 million cells per pellet C-14068
BPE-26 1 million cells per pellet C-14069
EGF, human, recombinant 1 million cells per pellet C-30020
C-30021
2 ml C-60170
2 ml
500 µg

PromoCell GmbH USA/Canada United Kingdom 11/2015
Phone: 1 – 866 – 251 – 2860 (toll free) Phone: 0800 – 96 03 33 (toll free)
Sickingenstr. 63/65 Fax: 1 – 866 – 827 – 9219 (toll free) Fax: 0800 – 169 85 54 (toll free)
69126 Heidelberg Other Countries
Germany Deutschland Phone: +49 6221 – 649 34 0
Telefon: 0800 – 776 66 23 (gebührenfrei) Fax: +49 6221 – 649 34 40
Email: [email protected] Fax: 0800 – 100 83 06 (gebührenfrei)
www.promocell.com © PromoCell GmbH 2015
France
Téléphone: 0800 90 93 32 (ligne verte)
Téléfax: 0800 90 27 36 (ligne verte)