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

“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:


                                                                  4
               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
                                                   0
                                          0
               vaccine increased by 0.8   to 1.2 C  at days 1 to 4 after the vaccination, and then it
               reached at normal level.





               Бүрэнмэндийн Баярцэцэг                                                                  50
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