Untuk melakukan kultur sel, hal yang harus dipersiapkan:

1. Laboratorium khusus yang terhindar dari kontaminasi apapun. Misalnya; mycoplasma,

bacteria, fungi,
2. Microbiological safety cabinet kelas 1 (untuk kultur cell line) (yang kayak di lab
biomedik)
3. Centrifuges
4. Inkubator. Sel kultur butuh lingkungan yang tepat untuk tumbuh dan berkembang.
Inkubator yang baik adalah yang memiliki pengaturan temperature, derajat humidity
dan CO2 pada keadaan stabil. Biasanya suhu yang dipakai untuk sel mamalia adalah
37C, dan kadar CO2 5-10%

Fase pertumbuhan sel:

1. Lag Phase at this stage the cells do not divide. During this period the cells
adapt to the culture conditions and the length of this phase will depend upon
the growth phase of the cell line at the time of subculture and also the seeding
density.
2. Logarithmic (Log) Growth Phase cells actively proliferate and an
exponential increase in cell density arises. The cell population is considered to
be the most viable at this phase, therefore it is recommended to assess cellular
function at this stage. Each cell line will show different cell proliferation kinetics
during the log phase and it is therefore the optimal phase for determining the
population doubling time. Cells are also generally passaged at late log phase.
Passaging cells too late, can lead to overcrowding, apoptosis and senescence.
3. Plateau (or Stationary) Phase cellular proliferation slows down due to the
cell population becoming confluent. It is at this stage the number of cells in the
active cell cycle drops to 0-10% and the cells are most susceptible to injury.
4. Decline Phase cell death predominates in this phase and there is a
reduction in the number of viable cells. Cell death is not due to the reduction
in nutrient supplements but the natural path of the cellular cycle.

1. Sanitise the cabinet using 70% isopropanol before commencing work.

2. Sanitise gloves by spraying them with 70% isopropanol and allowing to air
dry for 30 seconds before commencing work.
3. Put all materials and equipment into the cabinet prior to starting work.
Equipment in the cabinet or that which will be taken into the cabinet during
cell culture procedures (media bottles, pipette tip boxes, pipette aids) should
be wiped with tissue soaked with 70% isopropanol prior to use.
4. Whilst working do not contaminate gloves by touching anything outside the
cabinet (especially face and hair). If gloves become contaminated re-spray
with 70% isopropanol as above before proceeding.
5. Discard gloves after handling contaminated cultures and at the end of all cell
culture procedures.
6. Movement within and immediately outside the cabinet must not be rapid.
Slow movement will allow the air within the cabinet to circulate properly.
7. Speech, sneezing and coughing must be directed away from the cabinet so
as not to disrupt the airflows.
8. After completing work disinfect all equipment and material before removing
from the cabinet. Spray the work surfaces inside the cabinet with 70%
isopropanol and wipe dry with tissue. Dispose of tissue by incineration.
9. Liquid cell culture waste should be discarded in sodium hypochlorite
(10,000 ppm) and must be kept in the cabinet for a minimum of two hours
(preferably overnight) prior to discarding to the drain with copious amounts
of water.
10. Periodically clean the cabinet surfaces with a disinfectant or fumigate the
cabinet according to the manufacturers instructions. However, you must
ensure that it is safe to fumigate your own laboratory environment due to
the generation of gaseous formaldehyde, consult your on-site Health and
safety advisor