Thaum cov hlwb peb kab lis kev cai nyob rau hauv

cell Hoobkasmuaj kab mob, feem ntau ntawm lawv yog qhov nyuaj los daws.Yog tias cov kab mob sib kis muaj txiaj ntsig thiab nyuaj kom tau txais dua, cov txheej txheem hauv qab no tuaj yeem siv los tshem tawm lawv.

1. Siv tshuaj tua kab mob

Cov tshuaj tua kab mob zoo dua los tua cov kab mob hauvcell factories.Kev sib xyaw tshuaj muaj txiaj ntsig zoo dua li cov tshuaj ib leeg.Cov tshuaj tiv thaiv zoo dua li kev siv tshuaj tom qab kis kab mob.Cov tshuaj tiv thaiv feem ntau siv cov tshuaj tua kab mob ob npaug (penicillin 100u / mL ntxiv rau streptomycin 100μg / mL).Tom qab kev sib kis, txoj kev tu yuav tsum yog 5 mus rau 10 npaug ntau dua li qhov qub.Cov tshuaj yuav tsum tau siv rau 24 mus rau 48 teev tom qab ntxiv, thiab tom qab ntawd hloov nrog cov txheej txheem niaj hnub.Kab lis kev cai kua.Txoj kev no yuav ua tau zoo nyob rau theem pib ntawm kev sib kis.Ntxiv rau cov tshuaj penicillin thiab streptomycin, cov tshuaj tua kab mob siv tau kuj muaj xws li gentamicin, kanamycin, polymyxin, tetracycline, nystatin, thiab lwm yam. Feem ntau siv yog 400 mus rau 800 μg / mL kanamycin lossis 200 μg / mL tetracycline.Qhov nruab nrab yog hloov txhua 2 mus rau 3 hnub thiab dhau mus rau 1 mus rau 2 tiam rau kev kho mob.Hauv xyoo tas los no, nws tau tshaj tawm tias 4-fluoro, 2-hydroxyquinoline (Ciprofloxacin, Cip), Pleu-romutilin derivative (Pleu-romutilin derivative, BM-Cyclin2: BM-1 thiab tetracycline derivative (BM-2)) Cov tshuaj tua kab mob yog siv tau los tua mycoplasma thaum siv ib leeg lossis ua ke.Peb cov tshuaj tua kab mob no tau npaj tag nrho rau hauv 250X cov kev daws teeb meem hauv PBS thiab khaws cia ntawm -20 ° C rau kev siv tom qab.Kev siv cov concentration Cip yog 10 μg / mL, BM-1 yog 10 μg / mL, thiab BM-2 yog 5μg / mL.Thaum siv, thawj zaug aspirate cov kab lis kev cai nruab nrab, ntxiv RPMI1640 kab lis kev cai nruab nrab uas muaj BM-1, tom qab ntawd aspirate kab lis kev cai nruab nrab tom qab 3 hnub, ntxiv RPMI1640 kab lis kev cai nruab nrab uas muaj BM-2, thiab kab lis kev cai rau 4 hnub, thiab lwm yam rau 3 hnub sib law liag. .rounds, kom txog rau thaum nws tau ua pov thawj los ntawm 33258 fluorescent staining microscopy tias mycoplasma tau raug tshem tawm, ces ib txwm kab lis kev cai nruab nrab yog ntxiv rau kab lis kev cai thiab nqe lus 3-4 zaug.

2. Kev kho cua sov

Incubating kab mob kis kab mob ntawm 41 ° C rau 18 teev tuaj yeem tua cov mycoplasma, tab sis muaj kev cuam tshuam tsis zoo rau cov hlwb.Yog li ntawd, kev sim ua ntej yuav tsum tau ua ua ntej kev kho mob txhawm rau tshawb xyuas lub sijhawm cua sov uas tuaj yeem tua cov mycoplasma mus rau qhov siab tshaj plaws thiab muaj kev cuam tshuam tsawg tshaj plaws ntawm cov hlwb.Txoj kev no yog qee zaum tsis ntseeg.Yog tias kho nrog tshuaj ua ntej thiab tom qab ntawd kub ntawm 41 ° C, cov nyhuv yuav zoo dua.

3. Siv cov tshuaj mycoplasma tshwj xeeb

Cov kab mob mycoplasma tuaj yeem raug tshem tawm nrog 5% luav mycoplasma immune serum (hemagglutination titer 1: 320 lossis siab dua).Vim tias cov tshuaj tiv thaiv tshwj xeeb tuaj yeem cuam tshuam qhov kev loj hlob ntawm mycoplasma, nws hloov tsis zoo 11 hnub tom qab kev kho mob antiserum thiab tseem tsis zoo 5 lub hlis tom qab.yog qhov tsis zoo.Txawm li cas los xij, txoj kev no muaj teeb meem ntau dua thiab tsis yooj yim thiab pheej yig li siv tshuaj tua kab mob.

4. Lwm txoj kev

Ntxiv nrog rau cov txheej txheem saum toj no ntawm kev tshem tawm cov kab mob, kuj tseem muaj txoj hauv kev ua kom tsis muaj menyuam thiab ua kom tsis muaj menyuam hauv cov tsiaj, cov txheej txheem macrophage phagocytosis, txoj kev ntxiv bromouracil rau cov kab mob.lub raj mis kab lis kev caithiab tom qab ntawd irradiating lawv nrog lub teeb, thiab pom txoj kev, thiab lwm yam, tab sis lawv tag nrho cov teeb meem thiab tsis muaj txiaj ntsig.Yog li ntawd, thaum muaj kab mob mycoplasma tshwm sim, tshwj tsis yog nws muaj txiaj ntsig tshwj xeeb, nws feem ntau muab pov tseg thiab rov ua dua tshiab.

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