Escolar Documentos
Profissional Documentos
Cultura Documentos
Victoria Alonso
valonso@fbioyf.unr.edu.ar
Material de Vidrio
Tissue Culture
CELLSTAR® Cell Culture Products
1 Cell/
• Estéril por irradiación.
Cell Culture
• El plástico más empleado es el poli-estireno, Products
de buena calidad óptica. Debido a
Microplates
2 HTS-
que este plástico es hidrofóbico requiere un tratamiento
CELLSTAR ® con irradiación-gamma
Standard Cell Culture Flasks
o mediante descargas eléctricas que produzca una
For cell culture, superficie
Greiner hidrofílica.
Bio-One offers standard and filter The s
3 Immunology/
cap cell culture flasks with growth areas of 25 cm2, 75 cm2 to eff
• El tratamiento es característico de los diferentes proveedores, y por ello los
and 175 cm2. Suspension culture flasks complete the range provid
in the sizes of 50 ml, 250 ml, 550 ml and 650 ml. desig
productos varían en calidad de uno a otro.
HLA
All Greiner Bio-One cell culture flasks are made of high-grade mediu
polystyrene. For adherent cell culture, the surfaces of our conta
• Otros plásticos que se emplean son polivinilo-cloruro, policarbonato (PVC), poli
standard and filter cap cell culture flasks are physically ensur
4 Microbiology/
surface-treated to improve cell adhesion and proliferation. Both
Bacteriology
tetrafluoretileno (teflón, PTFE), etc.
Purpose Beakers
661 160
5 Tubes/Multi- 660 160
Sta
• Imp
658 170
thro
trea
• Ce
sta
Handling
6 Liquid
(wit
• Ca
• Gra
7 Molecular
690 160
Biology
690 170
Stand
n
Material de Plástico: Contenedores y superficies
• Sistema de cultivo
o Monocapa estacionaria è Flasks, placas
o Monocapa en movimiento è Botellas roller
o Suspensión estacionaria è Flasks o placas
o Suspensión en movimiento è Spinner flasks o bioreactores
Description
Triangular and modified Growth area (cm²)
Rectangular flasks haveWorking
a volume (mL)neck and Cell yield*RoboFlask vessels are
Angled
Triangular: buen acceso
triangular 35
de la pipeta
flasks offer rampyfromscraper
8 the bottom a lostoángulos. Su base
the 1 to 2traditional ancha
straight 0.8 ×provee mayor
106robotics-compatible c
estabilidad. good pipette60 and cell canted 21neck for easier pour- 4 to 5neck flasks utilize the 2.1 × 106culture flasks offering
Rectangular:scraper
pueden accesstener
100 to the una rampa desde el fondo del
ing and55pipette access. Most 10 to 12 cuello angular para
entire bottom area5.5 × 106facilitar
92.6 cmel
2
cell growth
volcado y el acceso
corners. Thea pipetas.
150 wider baseOtra alternativa
canted neck flasks
148 sonalsolas
have tradicionales con cuello
for cell growth.
28 to 32 14.8 ×angular
Their 106surface o
area. The flas
recto las cuales permiten
provides
*Cell que se upon
added stability.
line dependent. Based utilice toda
an antitip ofel1to×área
skirt
a density para el crecimiento
enhance
105 cells/cm². design saves oncelular.
space Su arediseño
designed for use
ahorra espacio y reduce el contacto del medio con el cuello yand
stability. la reduces
tapa. medium automated cell cultur
sloshing into the neck. systems utilizing a
recommended for use in all CO2 incubators, especially for
long-term use. The vent cap was a Corning innovation that
Forma de las botellas de cultivo
first appeared in 1988.
Cuellos
Flask2:26
5/17/2005 NeckPM Styles
Page 1
Angled
◗ Manufactured from optically clear virgin polystyrene
◗ Treated for optimal cell neck improves pipette access and reduces medium
attachment
Cuello
angular:
mejora
el
acceso
de
la
pipeta
y
reduce
el
contacto
del
sloshing
◗ Printed with lot numbers for easeinto the neck. This patented design was a Corning
in traceability
medio
con
la
tapa.
◗ 100% integrity tested innovation that first appeared in 1988.
Tapas
Certified nonpyrogenic
◗
gases.
Phenolic styleScaps
e
uare
san
en
sistemas
designed abiertos
(when loosened) que
requieren
la
entrada
de
gases.
for use in
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
µm
sellada
en
la
tapa
que
provee
un
intercambio
gaseoso
consistente
pero
Vent caps contain a 0.2 µm nonwettable membrane sealed
to the cap, providing consistent, sterile gas exchange while
minimizando
minimizing los
riesgos
the risk of contamination. These de
capscontaminación.
are highly Son
recomendadas
para
usar
recommended for use in all CO incubators, especially for
en
incubadores
long-term use. The vent capdwas
2
e
CaOCorning
2,
especialmente
innovation that para
uso
prolongado.
first appeared in 1988. 1
Like all Geiner Bio-One products, cell culture dishes are For exact dimensions of our cell culture dishes, please refer to the Culture Vessels and Surfa
manufactured according to our high quality standards. Dishes are product data sheets on our website.
Tissue Culture
Multiwell plates
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 CELLSTAR® Cell Culture Products
with a height of 20 mm is available. These widely used vessels were originally designed for virus titration, but have since become popu
1 Cell/
Placas con pocillos o multiwell 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
CELLSTAR® Cell Culture Multiwell Plates
number of dishes. They are more convenient to handle, especially if the pipettors, plate washers, rea
Microplates
and other equipment for processing these plates are used. They must be used with incubators that co
2 HTS-
662 102 humidity and CO2 levels.
Cell Culture Multiwell Plates
662 160
Cat.-No. 6, 627
12,160 627 170
24,Description
48 Well Format 628 160 632 171
Growth well (cm²) Working volume/ well (mL) Cell yield*
3 Immunology/
ø [mm] x height [mm] 35 x 10 35 x 10
96-well 60 x 15 0.32 94 x 16 0.1 to 0.2 0.32 × 105
Cell Culture Inserts p. 1 I 40 ff.
Growth area [cm2] 8.7 48-well
0.93/well 21 1.00 58 0.3 to 0.6 0.8 × 105
24-well 1.88 0.5 to 1.2 1.9 × 105
HLA
Total volume [ml] 665 102 10 10 28 80
665 180 12-well 3.83 1.0 to 2.4 3.8 × 105
Working volume [ml] 5 0.1/well 6–7 15 – 16
4 Microbiology/
657 160 6-well 9.40 2.0 to 3.0 9.5 × 105
Bacteriology
657 185 Cell culture multiwell plates are available in the following versions:
Vents + *Cell line dependent.
+ Based upon a density of 1 × 105 cells/cm².
+ -
• With hydrophilic surface • With hydrophobic surface
TC surface treatment + surface treatment)
(TC + for for+ suspension cultures
+
677 102 Roller bottles
improved cell adhesion and hybridoma cells
Sterile 677 180 + + + +
Purpose Beakers
The roller bottle was developed for cultivating large numbers of anchorage-dependent cells.²0 Today
5 Tubes/Multi-
Quantity per bag/case 10/740
provide 10/740
a more economical 10/600
means 10/480 large volumes of cells using essentially the same cu
for cultivating
Properties • Compatible with common instruments and automated systems.
techniques as with flasks
New but
For further information please
with considerably less labor. Besides the traditional smooth wall design,
visit our website.
• High clarity and low autofluorescence 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
numberBotellasof dishes.
"roller bottles".
roller
Se trata deThey
tubos, are more
con una cara convenient
plana sobre la queto
se handle, especially
fija el cultivo, if the
y que se incuban pipettors,
en los
Spining flask
incubadoresplate washers, readers,
◗
and other
dotados equipment
de "roller". for processing
Existen variantes
producción de gran número de células.
these plates
con una gran superficie are(espirales
de adhesión used. de They musty que
plástico...) be seused with
destinan a laincubators that control
◗
humidity and CO2 levels.
Description Growth well (cm²) Working volume/ well (mL) Cell yield* ◗
96-well 0.32 0.1 to 0.2 0.32 × 105
48-well 1.00 0.3 to 0.6 0.8 × 105 D
24-well 1.88 0.5 to 1.2 1.9 × 105 ◗
12-well 3.83 1.0 to 2.4 3.8 × 105
◗
6-well 9.40 2.0 to 3.0 9.5 × 105
*Cell line dependent. Based upon a density of 1 × 105 cells/cm². ◗
◗
Roller bottles
The roller bottle was developed for cultivating large numbers of anchorage-dependent cells.²0 Today they Ta
provide a more economical means for cultivating large volumes of cells using essentially the same culture
echniques as with flasks
Incubador but with considerably less labor. Besides
tipo “roller” the traditional smooth
“Roller bottles” Figure 11. Ready-to-use
wall design, roller disposable C
bottles are available
a fase gaseosa. withmássmall
Los componentes ridges
significativos de lathat approximately
fase gaseosa son el oxígeno ydouble the
el dióxido de surface area available
carbono: Corning forspinner
growingvessels avoid the 1
cells without
Oxígeno. increasing
Las necesidades thepara
de oxígeno dimensions
la mayor parteof de the bottles.
los cultivos celulares es cubierta con la tensión atmosférica, cleaning and sterilizing problems
2
aunque existen cultivos, especialmente los cultivos de órganos que requieren una tensión de oxígeno superior (del 95%)
posiblemente debido a la geometría del órgano y a las dificultades de difusión del gas en su interior. Se ha propuesto
common with reusable glass
(McKeehan y col.,Description
1976) que los requerimientosGrowth
de selenioarea
en el (cm²)
medio podrían serWorking
debidos avolume (mL)
su papel en la detoxificación Cellspinner
yield* flasks. 5
de radicales de oxígeno, especialmente en los medios sin proteínas séricas.
Small 490 100 to 150 4.9 × 107 1
Dióxido de carbono. El dióxido 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 8.5 × 107
3
Pharmaceutical 1750
⎯⎯→ ⎯⎯→
340 to 500 17.5 × 107
H2O + CO2 ←⎯⎯ H2CO3 ←⎯⎯ H+ + HCO3−
*Cell line dependent. Based upon a density of 1 × 105 cells/cm².
[1] 6
⎯⎯→
1
NaHCO3 Na+ + HCO3− [2]
Surface Coatings and Feeder Cells
←⎯⎯
3
El incremento de la concentración de ión bicarbonato desplaza la ecuación [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 ecuación: *A
Most tissue culture work uses disposable polystyrene †O
vessels. The vessel surface NaOH + H2COis 3 ←
treated
⎯ ⎯→ to render
⎯⎯→ it+
⎯⎯ NaHCO3 + H2O ←⎯⎯ Na + HCO3 + H2O
−
[3]
hydrophilic (wettable). Most cell lines in the ATCC
6
collection are cultivated on treated plastic surfaces in
De modo que para establecer un pH determinado se debe tener en cuenta especialmente los niveles de bicarbonato sódico
y la tensión de CO . Cada medio tiene una concentración recomendada de bicarbonato y tensión de CO para alcanzar el pH
2 2
correcto. Sin embargo, se emplean otras sustancias tamponadoras en la formulación de muchos medios en la actualidad, lo
Color-coded magnifier stripes make volume reading e
Grads. (mL)
Negative
10.0
10.0
N/A
0.2
0.2
2.5
3.0
3.0
0.2
2.5
0.2
0.2
Exclusive antidrip tip assures accurate delivery
Graduations
◗
Available in variety packs of white, Available in variety packs of white, screw cap
1/100
1/100
1/100
1/100
1/100
1/10
1/10
1/10
2/10
1/10
(mL)
Stripette® Serological Pipets
1/2
blue, green, red, and yellow. blue, green, red, and yellow.
1
◗ Inner cap ring
Silicone washer provides a secure Silicone washers or rubber assures a tight
n de células en placa
◗ Easy-to-read black graduations for Easy-to-read black graduations ◗
Capacity
partial volumes en las que es necesario separar las for partial volumes
(mL)
100
lar representan dos aplicaciones células entre sí y
10
10
25
50
1
1
5
2
2
d celular. Para conseguirlo es necesario romper la
◗ Self-standing base, self-locking skirtmalla de proteínas que forman la self-locking skirt
◗ Self-standing base,
2:29 PM
idas mediante procesos que separen las moléculas proteicas de la matriz extracelular
que permiten la unión), o bien rompiendo proteolíticamente (mediante la acción de
CLS4492*
CLS4011
CLS4012
CLS4051
CLS4101
CLS4251
CLS4501
CLS4484
CLS4485
CLS4487
CLS4021
CLS4486
Cat. No.
Warning! Do not use cryogenic vials for storage in the liquid phase of liquid nitrogen. Only store vials in the vapor phase above
pueden clasificar en tres categorías:
the liquified gas. Always use appropriate safety equipment when removing vials from cryogenic storage.
5/17/2005
Scrapers y pipetas
cribar, rascar, etc...). En cultivos celulares se emplean con frecuencia los métodos de
◗
◗
◗
◗
◗
◗
arrancar las células adheridas.
◗ Certified RNase-/DNase-free
◗ Vials have a silicone washer for a secure seal.
◗ Vials
a adición may been color
de soluciones las que coded with
no hay iones inserts
divalentes (Cat.
o bien No. 430499)
de agentes quelantes
educe la concentración de los iones que estabilizan las uniones de las proteínas de la
◗ Self-standing
eceptores celulares. vials have a special base design allowing them to be locked into cryogenic rack
Esterilización de material
• Colocar los materiales sin tocar el fondo del autoclave y que descansen
en una rejilla en posición vertical.
• No debe sobrecargarse el autoclave, evitar que la carga quede en contacto
con las paredes o con la puerta.
Polietileno 9 meses
Inverted
microscope
Refrigerator
Storage rack
Cell
Freezer counter Centrifuge
Water Prep
Bench with shelf over purifier bench
Sterilizing oven
Water
Sink bath
Liquid CO2 cylinders
with automatic Autoclave
changeover Deionised
water supply Air extract
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.
CULTURE OF ANIMAL CELLS
Service
PREPARATION cart MICROSCOPE
Water AREA Reagent shelving ROOM
purifier (sterile fluids)
Inverted Cell
microscope counter
Washup Air intake or
sinks Liquid location of air Two-tier
Storage nitrogen conditioner at incubator
rack storage high level
Water
Wash- Liquid CO2 Sink bath
up cart cylinders with
automatic Refrigerator
changeover Freezer
Storage CO2
rack incubators 37oC
ROOM
Washup cart Sink Centrifuge
Refrigerator
Freezer 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.