Page 50 - Хонь, ямааны цэцгийн вакцины сөөлжих идэвхийг сорьсон дүн

“Хонь, ямааны цэцгийн вакцины сөөлжих идэвхийг сорьсон дүн” 2015 он

Cell count technique

Nutrient medium of cell culture in plastic flask was removed and stuck cells were

peeled off withsolution of trypsin and EDTA (in 1:5 ratio).

1.Culture was transferred from plastic flask into tube and then centrifuged.

2.Two hundreds microliters of cell suspension from supernatant were taken
into 15 ml tube and 300 µl PBS and 500 µl methylen blue were added.

3.Twenty microliters of prepared suspension was taken and placed on
Goryaev’s chamber.Method of stained cell count in Goryaev’s chamber:Live

cells counts were determined in larger diagonal squares and then the counts

are estimatedby formula. Dead cells are stained in blue and live cells are clear
white color. The formula is as follows:

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Cell counts per 1 ml = cell counts x dilution x 10 Using above formula the cell counts
per 1 ml are calculated.

Calculation of the proportion of live cells

Formula 1Percentageof livecells= Live cell counts/Total cell counts х100

Live cell counts=5700/6500х 100%=87.62%

Cell storage:

Dulbecco’s nutrient medium was added, where DMSO and FCS accounted for 70%
and 20%respectively, and then the culture was preserved by shifting at temperature

gradients -40С, -200С,-800С, and -1960С. Before growing the preserved culture at
temperature 370С, nutrient mediumwas changed and adapted at temperature 370С

for 2 or 3 days and the culture is then passaged byconventional method.

Results

1. Body temperature of sheep and goats injected with sheep and goat pox virus
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vaccine increased by 0.8 to 1.2 C at days 1 to 4 after the vaccination, and then it
reached at normal level.

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