Material de Cultivo Celular

Las condiciones de cultivo varian ampliamente segn el tipo de clula utilizada,

pero el ambiente artificial donde las clulas se cutivan consiste invariablemente
de: un contenedor apropiado con un sustrato o medio que provea los
nutrientes esenciales (aminocidos, carbohidratos, vitaminas, minerales),
factores de crecimiento, hormonas y gases. La mayora de las clulas son
adherentes y deben ser cultivadas adheridas a un sustrato slido o semislido,
mientras que otras pueden crecer flotando en medio de cultivo (en suspencin).

Victoria Alonso
valonso@fbioyf.unr.edu.ar

Material de Vidrio
Empleado usualmente como sustrato de cultivo tiene como ventajas su escaso
costo y su facilidad de limpieza y esterilizacin. Asimismo es especialmente til
para su posterior observacin al microscopio por su calidad ptica. Actualmente
se utiliza material de plstico descartable para este fin.
Prcticamente todos los Medios se preparan y esterilizan para su
almacenamiento en botellas de vidrio.

Tambin se utilizan diferentes tipos de pipetas de cristal, probetas, vasos y

matraces para el trabajo rutinario del Laboratorio.

 Estril por irradiacin.

1 Cell/
Tissue Culture

Plstico descartable

CELLSTAR Cell Culture Products

2 HTSMicroplates

Cell Culture
Products
El plstico ms empleado es el poli-estireno,
de buena calidad
ptica. Debido a

que este plstico es hidrofbico requiere

un tratamiento
con irradiacin-gamma
CELLSTAR
Standard
Cell Culture Flasks

3 Immunology/
HLA

o mediante descargas elctricas que produzca

una
superficie
hidroflica.
For cell culture,
Greiner
Bio-One offers
standard and filter
cap cell culture flasks with growth areas of 25 cm2, 75 cm2
and 175 cm2. Suspension culture flasks complete the range
in the sizes of 50 ml, 250 ml, 550 ml and 650 ml.
All Greiner Bio-One cell culture flasks are made of high-grade
polystyrene. For adherent cell culture, the surfaces of our
standard and filter cap cell culture flasks are physically
surface-treated to improve cell adhesion and proliferation.

El tratamiento es caracterstico de los diferentes proveedores, y por ello los

productos varan en calidad de uno a otro.
4 Microbiology/
Bacteriology

Otros plsticos que se emplean son polivinilo-cloruro, policarbonato (PVC), poli

The s
to eff
provid
desig
mediu
conta
ensur
Both

660 160

661 160

Sta

Imp
thro
trea

658 170

Ce
sta
(wit

6 Liquid
Handling

5 Tubes/MultiPurpose Beakers

tetrafluoretileno (tefln, PTFE), etc.

7 Molecular
Biology

Ca

Gra
690 160
690 170

Stand

Material de Plstico: Contenedores y superficies

Se debe seleccionar segn los requerimientos de:
Caractersticas de las clulas:

Como elegir el adecuado??

o Anclaje dependiente
o Anclaje independiente
o Ambas
Sistema de cultivo
o Monocapa estacionaria Flasks, placas
o Monocapa en movimiento Botellas roller
o Suspensin estacionaria Flasks o placas
o Suspensin en movimiento Spinner flasks o bioreactores
Tamao del cultivo:
o 0,5-1 x 106 clulas confluentes/cm2
o 1-2 x 106 clulas/ ml de cultivo (en agitacin)

neck rectangular (also hexagonal) glass T-flasks in the 1940s. Today, plastic flasks are available with a range
of growing
areas, a variety of shapes, with several different neck designs. Choice of design depends on the
Botellas
o flasks
cell culture techniques used as well as personal preference. The more common sizes are listed below.
sigma.qxd 5/17/2005 2:27 PM Page 2

Tamaos

LT U R E

Description

Growth area (cm)

Cell yield*

25

Recommended working
volume (mL)
5 to 10

T-25
T-75
T-150

75
150

15 to 25
30 to 50

7.5 106
15.0 106

T-175

175

35 to 60

17.5 106

T-225

225

45 to 75

22.5 106

2.5 106

*Cell line dependent. Based upon a density of 1 105 cells/cm.

Flask Shapes

Cell culture
dishesa flask shape is usually a matter of personal preference:
Formas
Choosing
Cell culture dishes offer the best economy and access to the growth surface.
This makes them the vessels of choice for cloning or other manipulations such
as scraping that require direct access to the cell monolayer. They must be used
with incubators that control CO2 and humidity. Most manufacturers offer
dishes in four diameters: 35 mm, 60 mm, 100 mm, and 150 mm. These are
nominal diameters and may not be the actual diameter of the growth surface.
Cell culture dishes are available with either specially treated surfaces for
growing anchorage-dependent cells, or untreated (native) surfaces for
growing suspension cultures where attachment is not desired.

Description
Growth
area (cm)
volume
(mL)neck and Cell yield*RoboFlask vessels are
Angled
Rectangular
flasks haveWorking
a
Triangular
and modified
Triangular: buen
acceso
de la pipeta
scraper
a lostongulos.
Su base
ancha
mayor
8 the bottom
0.8 provee
106robotics-compatible
straight
c
rampyfrom
the 1 to 2traditional
triangular 35
flasks offer
estabilidad. good pipette
60 and cell
2.1 106culture flasks offering
canted 21
neck for easier pour- 4 to 5neck flasks utilize the
2
Rectangular:scraper
pueden
tener
una
rampa
desde
el
fondo
del
cuello
angular
para
facilitar
entire bottom area5.5 106
92.6 cmel
cell growth
ing and55pipette access. Most 10 to 12
100 to the
access
volcado y el acceso
a pipetas.
alternativa
son
tradicionales
con cuello
for cell growth.
Their
area. The flas
canted
neck flasks
alsolas
have
corners. The
wider baseOtra
150
148
28 to 32
14.8 angular
106surface o
design saves oncelular.
space Su
arediseo
designed for use
an antitip
skirt
enhance
provides
added stability.
recto las cuales
permiten
que
se upon
utilice
toda
para el crecimiento
*Cell
line dependent.
Based
a density
ofel1torea
105 cells/cm.
automated cell cultur
stability.
ahorra espacio y reduce el contacto
del medio con el cuello yand
la reduces
tapa. medium
sloshing into the neck.
systems utilizing a

Forma de las

recommended for use in all CO2 incubators, especially for

long-term use. The vent cap was a Corning innovation that
botellas
de cultivo
first appeared in 1988.

Cuellos
PM Styles
Page 1
Flask2:26
Neck

5/17/2005

Straight neck flasks are ideal for larger medium volumes

Cuello
recto: ideal para volmenes grandes de medio ya que disminuye
since this design reduces medium sloshing into the cap.
el contacto del medio con la tapa.
Canted neck flasks allow easier pouring and improved access
e Flasks
to the flask for pipetting or scraping. The canted neck
Cuello inclinado: permite volcar el contenido de la botella y hace ms
design waseal p
Corning
innovation
that first
sencillo
ipeteo
oof ssizes,
craping.
and appeared
Corning and Costar flasks
are
available
in
a
variety
designs
cap
in 1974.
styles to meet your needs.

Manufactured from optically clear virgin polystyrene
Angled
neck improves pipette access and reduces medium
Treated for optimal cell
attachment
Cuello angular: mejora el acceso de la pipeta y reduce el contacto del
sloshing
the neck. This patented design was a Corning
Printed with lot numbers
for easeinto
in traceability
100% integrity tested innovation that first appeared in 1988.
medio con la tapa.

Sterilized by gamma irradiation

Certified nonpyrogenic
Tapas

Flask Cap Styles

Tapas selladas: para usar en sistemas cerrados, no permiten el paso de

lquidos ni gases.

Tapas 2po fenlico: cuando se dejan ojas permiten el intercambio de
Phenolic
styleScaps
designed
(when loosened)
for use in
gases.
e uare
san
en sistemas
abiertos
que requieren la entrada de gases.
open systems requiring gas exchange. With the caps slightly

loosened,
gas is exchanged between the environments
inside and outside of the flask.
Tapas ven2ladas: con?enen una membrana impermeable de 0.2 m
Vent caps contain a 0.2 m nonwettable membrane sealed
sellada en la tapa que provee un intercambio gaseoso consistente pero
to the cap, providing consistent, sterile gas exchange while
minimizing
the risk of contamination.
These
caps
are highly
minimizando
los riesgos
de
contaminacin.
Son recomendadas para usar
recommended for use in all CO incubators, especially for
long-term
use. The vent capdwas
innovation that
en incubadores
e CaOCorning
para uso prolongado.
2, especialmente
Plug seal caps feature one-piece linerless construction and
are designed for use in closed systems, providing a liquidand gas-tight seal. When loosened, this cap can also be
used in open systems. This cap design was a Corning innovation that first appeared in 1974.

first appeared in 1988.

Flask Neck Styles

Straight neck flasks are ideal for larger medium volumes

with incubators that control CO2 and humidity. Most manufacturers offer
dishes in four diameters: 35 mm, 60 mm, 100 mm, and 150 mm. These are
nominal diameters and may not be the actual diameter of the growth surface.
Cell culture dishes are available with either specially treated surfaces for
growing anchorage-dependent cells, or untreated (native) surfaces for
Cell suspension
Culture cultures
Disheswhere attachment is not desired.
growing

Placas Cell Culture Dishes

CELLSTAR
628 160

Improved Description
cell adhesion
through physical surface
35
treatment

2
8.7 Growth
to 143 cm
area (cm)
growth areas

Working volume (mL)

8
1 to 2
Easy stacking
60
21
4 to 5
Vents ensure optimal
55
10 to 12
Maximal transparency
gas exchange 100
for excellent microscopic
150
148
28 to 32
analysis
Sterile and user-friendly
packaging
*Cell line dependent. Based upon a density of 1 105 cells/cm.

664 160
627 160

Available in the sizes

35, 60, 94, 100 and
page 21
145 mm

Like all Geiner Bio-One products, cell culture dishes are

manufactured according to our high quality standards. Dishes are
available in a wide variety of different dimensions and growth areas.
In the case of 58 cm2 and 143 cm2 dishes, an extra high profile
with a height of 20 mm is available.

New: 35 dish also

available with 4 internal
wells
www.atcc.org

For exact dimensions of our cell culture dishes, please refer to the
product data sheets on our website.

627 170
160
632 171
6, 627
12,160
24,Description
48 Well
Format 628
Growth well (cm)
Working volume/ well (mL)

Cell yield*
0.32 105

1.00 58
1.88

0.3 to 0.6
0.5 to 1.2

0.8 105
1.9 105

12-well
3.83
1.0 to 2.4
5
0.1/well
67
15 16
6-well
9.40
2.0 to 3.0
Cell culture multiwell plates are available in the following versions:
+ *Cell line dependent.
+
+
Based upon a density of 1 105 cells/cm.

3.8 105
9.5 105

657 160
657 185

TC surface treatment
677 102
Sterile
677 180
Quantity per bag/case
Properties

High clarity and low autofluorescence

10

24-well
10

With hydrophilic surface

+ surface treatment)
+
(TC
for
Roller
bottles
improved
cell adhesion
+
+

28

80

With hydrophobic surface

+
for+ suspension cultures
and hybridoma cells
+
+

4 Microbiology/
Bacteriology

665 102
665 180

Total volume [ml]

3 Immunology/
HLA

0.1 to 0.2

5 Tubes/MultiPurpose Beakers

Growth area [cm2]

Vents

0.32 94 x 16

35 x 10
35 x 10
60 x 15
96-well
Cell Culture Inserts p. 1 I 40 ff.
48-well
8.7
0.93/well
21

[mm] x height [mm]

Working volume [ml]

Culture Vessels and Surfa

2 HTSMicroplates

662 102
662 160
Cat.-No.

Email tech@atcc

Multiwell plates
CELLSTAR Cell Culture Products
These widely used vessels were originally designed for virus titration, but have since become popu
many other applications, especially hybridoma production, high-throughput screening, and tox
testing. Multiwell plates offer significant savings in space, media, and reagents when compared to an e
Plates
number of dishes. They are more convenient to handle, especially if the pipettors, plate washers, rea
and other equipment for processing these plates are used. They must be used with incubators that co
humidity
and CO2 levels.
Cell Culture
Multiwell Plates

Placas con pocillos o multiwell

CELLSTAR Cell Culture Multiwell

Cell yield*
0.8 106
2.1 106
5.5 106
14.8 106

1 Cell/
Tissue Culture

639 160

The roller bottle was developed for cultivating large numbers of anchorage-dependent cells.0 Today
10/740
10/740
10/600
10/480 large volumes of cells using essentially the same cu
provide
a more economical
means
for cultivating
Compatible with common instruments and automated systems.
techniques
as with flasks
with considerably less labor. Besides the traditional smooth wall design,
New but
For further information please
visit our website.
bottles are available with small ridges that approximately double the surface area available for gro
Easy-to-open packaging

in a wide range of sizes including

many other applications, especially hybridoma production, high-throughput screening, and toxicity
the 250 mL and 1L shown here.
esting. Multiwell plates offer significant savings in space, media, and reagents when compared to an equal
Botellas
roller
Spining flask
number
of dishes.
more
especially
if the
pipettors,
"roller bottles".
Se trata deThey
tubos, are
con una
cara convenient
plana sobre la queto
se handle,
fija el cultivo,
y que se incuban
en los
incubadoresplate washers, readers,
dotados
de "roller".
Existen variantes
con una gran superficie
de adhesin
plstico...)
destinan
a laincubators that control
and
other
equipment
for processing
these plates
are(espirales
used. de
They
musty que
be seused
with
produccin de gran nmero de clulas.
humidity and CO2 levels.
Description

Growth well (cm)

Working volume/ well (mL)

96-well
0.32
0.1 to 0.2
48-well
1.00
0.3 to 0.6
24-well
1.88
0.5 to 1.2
12-well
3.83
1.0 to 2.4
6-well
9.40
2.0 to 3.0
*Cell line dependent. Based upon a density of 1 105 cells/cm.

Cell yield*
0.32 105
0.8 105
1.9 105
3.8 105
9.5 105

Small

490

100 to 150

Dixido de carbono. El dixido de carbono juega un complejo papel en el medio debido a que influye la cantidad de CO2
disuelto, el pH y la Standard
cantidad de iones HCO3-. Las reacciones850
que tienen lugar en el medio 170
son: to 250

Pharmaceutical
1750
340 to 500

H2O + CO2 H2CO3 H+ + HCO3

*Cell line dependent. Based upon a density of 1 105 cells/cm.
NaHCO3

4.9 107
8.5 107
17.5 107

[1]

Na+ + HCO3

[2]

Surface Coatings and Feeder Cells

El incremento de la concentracin de in bicarbonato desplaza la ecuacin [1] hacia la izquierda, de modo que el pH se
establezca en 7,4. Se puede emplear asimismo otra base, por ejemplo NaOH, siendo la ecuacin:

Most tissue culture work uses disposable polystyrene

it+

vessels. The vessel surface

is 3
treated
to render
NaOH + H2CO
NaHCO3 + H2O Na + HCO3 + H2O
hydrophilic (wettable). Most cell lines in the ATCC
De modo que para establecer un pH determinado se debe tener en cuenta especialmente los niveles de bicarbonato sdico
collection
are cultivated on treated plastic surfaces in
y la tensin de CO . Cada medio tiene una concentracin recomendada de bicarbonato y tensin de CO para alcanzar el pH
2

Roller bottles
The roller bottle was developed for cultivating large numbers of anchorage-dependent cells.0 Today they
provide a more economical means for cultivating large volumes of cells using essentially the same culture
echniques as
with flasks
but with considerably less labor. Besides
the traditional smooth
wall design,
roller
Figure
11. Ready-to-use
disposable
Incubador
tipo roller
Roller bottles
bottles
are available
withmssmall
ridges
approximately
the
surface area available
forspinner
growingvessels avoid the
Corning
a fase gaseosa.
Los componentes
significativos
de lathat
fase gaseosa
son el oxgeno ydouble
el dixido de
carbono:
cells
without
increasing
thepara
dimensions
bottles.
cleaning and sterilizing problems
Oxgeno.
Las necesidades
de oxgeno
la mayor parteof
de the
los cultivos
celulares es cubierta con la tensin atmosfrica,
aunque existen cultivos, especialmente los cultivos de rganos que requieren una tensin de oxgeno superior (del 95%)
common with reusable glass
posiblemente debido a la geometra del rgano y a las dificultades de difusin del gas en su interior. Se ha propuesto
Working
(mL)
yield* flasks.
(McKeehan y col.,Description
1976) que los requerimientosGrowth
de selenioarea
en el (cm)
medio podran ser
debidos avolume
su papel en
la detoxificacin Cellspinner
de radicales de oxgeno, especialmente en los medios sin protenas sricas.

correcto. Sin embargo, se emplean otras sustancias tamponadoras en la formulacin de muchos medios en la actualidad, lo

Ta

1
2
5
1
3
6
1
3

*A
O
[3]

Exclusive antidrip tip assures accurate delivery

Available in 25, 50, and 100 mL sizes
Color-coded magnifier stripes make volume reading e
Bidirectional graduations provide choice of ascending
Negative graduations allow additional working volum
Three packaging options:
- Individually wrapped, clear plastic
- Individually wrapped, paper/plastic
- Bulk packed for large-scale sterile and nonsterile li

Stripette Pipets Ordering Information

n de clulas
en placa
Easy-to-read black graduations for

Graduations
(mL)

Silicone washer provides a secure

seal.

Silicone washers or rubber

O-rings provide a secure seal.
Capacity
(mL)

idas mediante procesos que separen las molculas proteicas de la matriz extracelular
que permiten la unin), o bien rompiendo proteolticamente (mediante la accin de

Cat. No.

2:29 PM

0.2
0.2
0.2
2.5
3.0
3.0
10.0
10.0
N/A

Easy-to-read black graduations

partial
volumes en las que es necesario separar las for
partial
volumes
lar representan dos
aplicaciones
clulas
entre
s y
d celular. Para conseguirlo
es
necesario
romper
la
malla
de
protenas
que
forman
la self-locking skirt
Self-standing base, self-locking skirt
Self-standing base,

External Thread
Plug Seal Cap

Individually Wrapped, Clear Plastic Wrap

CLS4011
1
1/100
CLS4012
1
1/100
CLS4021
2
1/100
CLS4051
5
1/10
CLS4101
10
1/10
CLS4492*
10
1/10
CLS4251
25
2/10
CLS4501
50
1/2
CLS4484
100
1

Page 23

Color-coded polypropylene
cap
Color-coded polypropylene cap
www.cultek.com
inserts simplify vial identification.
inserts simplify vial identification.
Available in variety packs of white,
Available in variety packs of white,
blue, green, red, and yellow.
blue, green, red, and yellow.

Stripette Serological Pipets

Internal Thread Cryogenic Vial

Individually Wrapped, Paper/Plastic Wrap

CLS4485
1
1/100
0.2
CLS4486
2
1/100
0.2
CLS4487
5
1/10
2.5

External Thread Cryogenic Vial

Negative
Grads. (mL)

Corning offers three styles of cryogenic vials as well as storage racks and boxes.
Crioviales

Sure-grip plug seal

screw cap

Inner cap ring

assures a tight
seal.

Warning! Do not use cryogenic vials for storage in the liquid phase of liquid nitrogen. Only store vials in the vapor phase above
the liquified gas. Always use appropriate safety equipment when removing vials from cryogenic storage.

5/17/2005

pueden clasificar en tres categoras:

Scrapers y pipetas

cribar, rascar, etc...). En cultivos celulares se emplean con frecuencia los mtodos de
arrancar las clulas adheridas.

Pipets

C E L L C U LT U R E

Stripette Serological Pipets

2 Cell Culture PSG_sigma.qxd

Manufactured from polypropylene to withstand temperatures down to -196C

Larger marking spot
Black graduations
Certified RNase-/DNase-free
Vials have a silicone washer for a secure seal.
a adicin
de soluciones
las que coded
no hay iones
divalentes
o bien
de agentes
quelantes
Vials
may been color
with
inserts
(Cat.
No. 430499)
educe la concentracin de los iones que estabilizan las uniones de las protenas de la
eceptores
celulares.
Self-standing
vials have a special base design allowing them to be locked into cryogenic rack

Three packaging options

External Thread Cryogenic Vials

Esterilizacin de material
La necesidad de asepsia para el cultivo celular se extiende no slo al lugar en que
se realiza el trabajo sino muy especialmente a los recipientes en que se realiza el
cultivo, a los medios lquidos o slidos, y a los instrumentos que puedan
entrar en contacto con ste en algn momento de su manipulacin (pipetas,
puntas de pipeta automtica, pinzas, tubos, material variado de vidrio...). Para
esterilizar todo este material, de variada naturaleza se emplean una serie de
mtodos: irradiacin con radiacin gamma o rayos X, esterilizacin por gas,
autoclavado, filtracin,... En el laboratorio general de cultivo de tejidos se suele
disponer de: equipo de filtracin y autoclave.

La esterilizacin por gas, y la irradiacin son tcnicas propias de grandes

instalaciones, por ejemplo hospitalarias o industriales

Equipos de filtracin. En la actualidad se dispone de unidades de filtracin estril muy recomendables para el trabajo
rutinario. Sin embargo, su elevado coste por unidad hace que en muchos casos an se empleen unidades reutilizables.
Estos equipos de filtracin constan o bien de una fuente de vaco conectada a un kitasato dotado de una unidad de
filtracin esterilizada por autoclavado, o bien de una bomba peristltica que fuerza el flujo de solucin a travs de una
unidad de filtracin hermtica. En ambos casos la esterilizacin se produce al atravesar la solucin un filtro de poro 0.22
m.

Equipos de filtracin. En la actualidad se dispone de

unidades de filtracin estril muy recomendables para el
trabajo rutinario. Sin embargo, su elevado costo por
unidad hace que en muchos casos an se empleen
unidades reutilizables. Estos equipos de filtracin
constan o bien de una fuente de vaco conectada a un
kitasato dotado de una unidad de filtracin esterilizada
por autoclavado, o bien de una bomba peristltica que
fuerza el flujo de solucin a travs de una unidad de
filtracin hermtica. En ambos casos la esterilizacin se
Autoclave.
autoclave es unun
instrumento
la esterilizacin
produce al atravesar
laUnsolucin
filtro que
depermite
poro
0.22 por calor tanto de slidos como de lquidos. La
esterilizacin se realiza habitualmente a 121C, 1 atmsfera de sobrepresin durante un tiempo superior a 20 min. Un
m.
autoclave de uso normal en el laboratorio de cultivo de tejidos suele disponer de temporizador y regulador de presin, y en
algunos casos de un ciclo de secado para permitir secar el material slido (material de vidrio, instrumentos quirrgicos,
tubos,...).

Autoclave. Un autoclave es un instrumento que permite la esterilizacin por calor

tanto de slidos como de lquidos. La esterilizacin se realiza habitualmente a
121C, 1 atmsfera de sobrepresin durante un tiempo superior a 20 min. Un
autoclave de uso normal en el laboratorio de cultivo de tejidos suele disponer de
temporizador y regulador de presin, y en algunos casos de un ciclo de secado
para permitir secar el material slido (material de vidrio, instrumentos quirrgicos,
tubos,...).

NORMAS PARA ESTERILIZAR EN AUTOCLAVE

Las dimensiones de un paquete deben ser como mximo de 30X30X50
cm. Y su peso no superior a 5 kg.
No deben ser hechos en forma muy compacta, es decir apretada para
que pueda penetrar y vapor con facilidad.
.
Se deben colocar controles de esterilizacin en el centro y exterior de
cada paquete a esterilizar.
Colocar los paquetes en posicin vertical.
Dejar espacios entre los paquetes.
Colocar los materiales sin tocar el fondo del autoclave y que descansen
en una rejilla en posicin vertical.

 No debe sobrecargarse el autoclave, evitar que la carga quede en contacto

con las paredes o con la puerta.
Los frascos o botellas, se deben colocar abiertos en posicin para permitir la
salida del aire.
Esterilizar cada tipo de material por separado. Materiales porosos con
porosos y no porosos con no porosos.
Cronometrar el ciclo, slo despus que el termmetro de la cmara cargada,
marque la temperatura deseada ( 121 C) previa verificacin de la presin.
Al abrir el autoclave conviene dejarlo 5 minutos hasta que escapen los
vapores
Controlar peridicamente con cultivos especiales de esporas
Despus de cada reparacin se debe controlar con cultivos bacteriolgicos.

DURACIN ENVOLTURA ESTANTE CERRADO

Papel kraft envoltura nica

3 semanas

Polietileno

9 meses

Empaques plsticos y sellados

al calor

1 ao

Tcnicas de manipulacin estril

La caracterstica principal que define al laboratorio de cultivo celular es el
mantenimiento de la asepsia. La tasa de crecimiento de las clulas en
cultivo es muy inferior al de los contaminantes habituales: hongos,
levaduras, bacterias y micoplasmas, y por ello para el mantenimiento del
cultivo ser vital evitar la aparicin en ste de cualquier
microorganismo indeseado.

Diagrama de la sala de cultivo

CHAPTER 4 LABORATORY DESIGN AND LAYOUT

Regular incubator

Air intake or location of

air conditioner

Humid CO2 incubator

Laminar-flow hood
(class II biological
safety cabinet) with
total extract
Service cart

CO2 supply

Inverted
microscope

Refrigerator
Freezer

Cell
counter

Bench with shelf over

Water
purifier

Storage rack
Centrifuge
Prep
bench
Sterilizing oven

Sink
Liquid CO2 cylinders
with automatic
changeover

Water
bath

Deionised
water supply

Main entry: double

asymmetric doors

1m

Autoclave
Air extract
Two-tiered high- &
low-temperature
drying oven

Soaking sinks Washing sinks

Fig. 4.1. Small Tissue Culture Laboratory. Suggested layout for simple, self-contained tissue culture
laboratory for use by two or three persons. Dark-shaded areas represent movable equipment,
lighter-shaded areas fixed or movable furniture. Scale 1:100.

45

CULTURE OF ANIMAL CELLS

Bench-top
autoclave

Sterilizing and
drying oven

Laminar-flow (biohazard) cabinets with total extract

PREPARATION
AREA

Water
purifier

Service
cart
Reagent shelving
(sterile fluids)
Inverted
microscope

Washup
sinks
Storage
rack
Washup cart

Liquid
nitrogen
storage
Liquid CO2
cylinders with
automatic
changeover

Cell
counter

Air intake or
location of air
conditioner at
high level

Two-tier
incubator
Water
bath

Sink
Refrigerator
Freezer

CO2
incubators

Storage
rack
Washup cart

Refrigerator
Freezer

MICROSCOPE
ROOM

Sink

37oC
ROOM

Centrifuge

CO2
Supply

Soaking
bath

1m
Main entry: double
asymmetric doors

Collection
tray

Air extract

Cryofreezer
(below)

CO2 Supply

Fig. 4.3. Tissue Culture Lab with Adjacent Prep Room. Medium-sized tissue culture lab (see Fig. 4.2),
but with attached preparation area, microscope room. and 37 C room. Scale 1:100.

Air inlet (extracts via hoods)

rea de trabajo dentro de la cabina de bioseguridad