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Kodi nchifukwa ninji chikhalidwe cha ma cell chimasokoneza maselo ofiira a magazi poyamba?

2022-12-23

Mawu oyamba
Erythrocyte lysate ndi imodzi mwa njira zosavuta komanso zosavuta kuchotsa maselo ofiira a magazi, ndiko kuti, kugawaniza maselo ofiira a magazi ndi lysate, omwe sawononga maselo a nucleated ndipo amatha kuchotsa kwathunthu maselo ofiira a magazi. Lysate cleavage ndi njira yochepetsera yochotsa maselo ofiira a m'magazi, yomwe imagwiritsidwa ntchito makamaka pakulekanitsa ndi kuyeretsa ma cell a minofu omwe amabalalitsidwa ndi digestion ya enzyme, kupatukana ndi kuyeretsedwa kwa ma lymphocytes, ndikuchotsa maselo ofiira amagazi pakuyesa mapuloteni a minofu ndi nucleic. kuchotsa asidi. Maselo a minofu omwe amapangidwa ndi lysate ya maselo ofiira a magazi alibe maselo ofiira a magazi, ndipo amatha kugwiritsidwa ntchito pa chikhalidwe choyambirira, kusakanikirana kwa maselo, kutuluka kwa cytometry, kulekana ndi kuchotsa nucleic acid ndi mapuloteni, etc.

Malangizo ogwiritsira ntchito
Chitsanzo cha maselo a minofu
1. Minofu yatsopano idagayidwa ndi kapamba / enzyme kapena collagenase ndipo imamwazikana mu kuyimitsidwa kwa selo limodzi, ndipo supernatant idatayidwa ndi centrifugation.

2. Tengani ELS lysate kuchokera mufiriji pa 4â, onjezerani ELS lysate ku cell precipitate mu chiŵerengero cha 1: 3-5 (kuwonjezera 3-5ml wa lysate ku 1ml wa selo lopangidwa), mokoma kuwomba ndi kusakaniza.

3. Centrifuge pa 800-1000rpm kwa mphindi 5-8 ndikutaya madzi ofiira owoneka bwino.

4. Gawo lomwe linasokonekera linasonkhanitsidwa ndikukhazikika ndi Hank's solution kapena serum-free culture solution kwa nthawi 2-3.

5, ngati kusweka sikokwanira/kutha kubwerezedwa masitepe 2 ndi 3.

6. Maselo obwezeretsanso zoyesera zotsatila; Ngati RNA yachotsedwa, ndi bwino kutero mu yankho lokonzedwa kuchokera ku Gawo 4 pogwiritsa ntchito madzi a DEPC

Maselo ofiira a magazi amakhala ndi moyo wautali kwambiri, masiku 120 okha, koma amabereka magazi mofulumira kwambiri, ndipo pamenepa amatha kugawanitsa maselo, ndipo ndi maselo omwe amagawanitsa mofulumira kuposa onse, kotero selo ili ndilofunika kwambiri. kotero ndizothandiza kwambiri pama cell chikhalidwe. Ndizosavuta, zilibe organelles mkati mwake, ma cell membranes ndi mapuloteni.

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