Chidule: Mesenchymal stem cell ndi kagawo kakang'ono ka pluripotent stem cell omwe amatha kudzipatula ku mesoderm.Ndi mawonekedwe awo odzibwereza okha komanso kusiyanitsa njira zambiri, ali ndi kuthekera kwakukulu kwamankhwala osiyanasiyana azachipatala.Maselo a mesenchymal stem ali ndi phenotype yapadera ya chitetezo chamthupi komanso mphamvu ya chitetezo chamthupi.Chifukwa chake, ma cell stem cell a mesenchymal amagwiritsidwa ntchito kale kwambiri pakuyika ma cell cell, uinjiniya wa minofu, ndi kuyika ziwalo.Ndipo kupitilira izi, amagwiritsidwa ntchito ngati chida choyenera muumisiri wa minofu ngati ma cell a seeder pamayesero oyambira komanso azachipatala.Mpaka pano, palibe njira yovomerezeka komanso yovomerezeka yoyendetsera bwino ma cell tsinde a mesenchymal.The Countstar Rigel imatha kuyang'anira kukhazikika, kuthekera, ndi mawonekedwe a phenotype (ndi kusintha kwawo) panthawi yopanga ndi kusiyanitsa ma cell tsindewa.The Countstar Rigel ilinso ndi mwayi wopeza zidziwitso zowonjezera za morphological, zoperekedwa ndi zojambulira zosatha za brightfield ndi fluorescence-based image pa nthawi yonse yowunikira khalidwe la cell.The Countstar Rigel imapereka njira yachangu, yotsogola, komanso yodalirika yowongolera ma cell tsinde.
Zida ndi njira:
Adipose-derived mesenchymal stem cell (AdMSCs) adapatsidwa mphatso ndi Pulofesa Nianmin Qi, AO/PI staining solution (Shanghai RuiYu, CF002).Antibody: CD29, CD34, CD45, CD56, CD73, CD105, HLADR (BD Company).
Ma AdMSC adapangidwa mu chofungatira cha 37 ℃, 5% CO2.Idyani ndi trypsin musanagwiritse ntchito.
Njira yodetsa ma CD idatsatiridwa ngati buku la antibody.
Kuzindikira kwa chikhomo cha CD ndi Countstar Rigel:
1. Njira yogwiritsira ntchito mtundu wa chizindikiro idapangidwa pokhazikitsa njira ya PE kukhala chithunzi cha PE fluorescence.
2. Minda itatu idalandidwa mchipinda chilichonse.
3. Pambuyo pa kujambula ndi kusanthula koyambirira kumalizidwa, malo olowera (log gate) kuti atenge kachilombo koyambitsa matenda ndi koyipa adakhazikitsidwa ndi mapulogalamu a FCS.
Kuwongolera kwamtundu wa stem cell
Chithunzi chotsatira (Chithunzi 1) chikuwonetsa ndondomeko ya stem cell therapy .
Chithunzi 1: Ndondomeko ya stem cell therapy
Zotsatira:
Kuzindikira kukhazikika, kuthekera, kukula, ndi kuphatikiza kwa ma AdMSC.
Kuthekera kwa ma AdMSC kunatsimikiziridwa ndi AO/PI, Njira yogwiritsira ntchito mitundu iwiri idapangidwa pokhazikitsa njira yobiriwira ndi kanjira Yofiyira ku chithunzi cha AO ndi PI fluorescence, kuphatikiza gawo lowala.Zithunzi zachitsanzo zawonetsedwa mu Chithunzi 2.
Chithunzi 2. Zithunzi za ma AdMSC asananyamuke komanso pambuyo pake.A. Asananyamuke;chithunzi choyimira chikuwonetsedwa.B. Pambuyo pa mayendedwe;chithunzi choyimira chikuwonetsedwa.
Kuthekera kwa ma AdMSC kudasinthidwa kwambiri pambuyo pa mayendedwe poyerekeza ndi zoyendera zisanachitike.Kuthekera kusanachitike mayendedwe kunali 92%, koma kudatsika mpaka 71% pambuyo pamayendedwe.Chotsatira chawonetsedwa pa Chithunzi 3.
Chithunzi 3. Zotsatira za kukhalapo kwa ma AdMSC (Asananyamuke komanso pambuyo pa mayendedwe)
M'mimba mwake ndi kuphatikiza zidatsimikiziridwanso ndi Countstar Rigel.Makulidwe a ma AdMSC adasinthidwa kwambiri pambuyo pa mayendedwe poyerekeza ndi asanayendetse.Kuzama kwake kusanachitike mayendedwe kunali 19µm, koma kudakwera mpaka 21µm pambuyo pa mayendedwe.Kuphatikizika kwa zisanachitike zoyendera kunali 20%, koma kudakwera mpaka 25% pambuyo pamayendedwe.Kuchokera pazithunzi zojambulidwa ndi Countstar Rigel, phenotype ya AdMSCs idasinthidwa kwambiri pambuyo poyenda.Zotsatira zawonetsedwa mu Chithunzi 4.
Chithunzi 4: Zotsatira zake ndi kukula kwake.A: Zithunzi zoyimira za AdMSCs, phenotype ya AdMSCs inasinthidwa kwambiri pambuyo pa mayendedwe.B: Kuphatikizikako kusanachitike mayendedwe kunali 20%, koma kudakwera mpaka 25% pambuyo pa mayendedwe.C: Kutalika kwake kusananyamuke kunali 19µm, koma kudakwera mpaka 21µm pambuyo pa mayendedwe.
Dziwani za immunophenotype ya AdMSCs yolembedwa ndi Countstar Rigel
Ma immunophenotype a AdMSC adatsimikiziridwa ndi Countstar Rigel, AdMSCs adalumikizidwa ndi ma antibody osiyanasiyana motsatana (CD29, CD34, CD45, CD56, CD73, CD105, HLA-DR).Njira yogwiritsira ntchito mtundu wa chizindikiro idapangidwa pokhazikitsa njira Yobiriwira kuti iwonetse PE fluorescence, kuphatikiza gawo lowala.Gawo lofotokozera zazithunzi zowala linagwiritsidwa ntchito ngati chigoba kuti muyese chizindikiro cha PE fluorescence.Zotsatira za CD105 zidawonetsedwa (Chithunzi 5).
Chithunzi 5: Zotsatira za CD105 za AdMSC zinatsimikiziridwa ndi Countstar Rigel.A: Kusanthula kwachulukidwe kwa kuchuluka kwa CD105 mu zitsanzo zosiyanasiyana ndi pulogalamu ya FCS Express 5 kuphatikiza.B: Zithunzi zapamwamba zimapereka chidziwitso chowonjezera cha morphological.C: Zotsatira zovomerezeka ndi tizithunzi za selo iliyonse, zida za pulogalamu ya FCS zidagawa ma cell m'magulu osiyanasiyana malinga ndi mawonekedwe awo a protein.
Zotsatira zina za ma antibodies zikuwonetsedwa mu Mkuyu 6
Chithunzi 6: A: Chithunzi choyimira ma ASC okhala ndi mawonekedwe owoneka ngati spindle.Kutengedwa ndi microscope ya OLYMPUS.Kukulitsa koyambirira, (10x).B: Kusiyana kwa Adipogenic kwa ASCs kumatsimikiziridwa ndi Ruthenium Red staining kusonyeza madera a mineralization.Kutengedwa ndi microscope ya OLYMPUS.Kukulitsa koyambirira (10x).C: Mawonekedwe a Countstar FL a ASC.
Chidule cha nkhani:
The Countstar FL imatha kuyang'anira kukhazikika, kuthekera, ndi mawonekedwe a phenotype (ndi kusintha kwawo) panthawi yopanga ndi kusiyanitsa ma cell tsindewa.FCS Express imapereka ntchitoyi kuti iwunikenso selo lililonse lazizindikiro, kutsimikizira deta kudzera mu chithunzi.Wogwiritsanso ntchito amathanso kukhala ndi chidaliro kuti achite zoyeserera zotsatila kutengera zotsatira za Countstar Rigel.The Countstar Rigel imapereka njira yachangu, yotsogola, komanso yodalirika yowongolera ma cell tsinde.
