Puhi: ʻO nā pūnaewele mesenchymal kahi ʻāpana o nā pūnaewele pluripotent hiki ke hoʻokaʻawale ʻia mai ka mesoderm.Me kā lākou hoʻololi hou ʻana a me nā hiʻohiʻona ʻokoʻa ʻokoʻa, loaʻa iā lākou ka mana kiʻekiʻe no nā ʻano lapaʻau like ʻole.Loaʻa i nā cell stem Mesenchymal kahi phenotype immune kūʻokoʻa a me ka hiki ke hoʻoponopono pale.No laila, ua hoʻohana nui ʻia nā cell stem mesenchymal i ka hoʻololi ʻana i nā cell cell, ʻenekinia kiko, a me ka hoʻololi ʻana o ke kino.A ma waho aʻe o kēia mau noi, ua hoʻohana ʻia lākou ma ke ʻano he mea hana kūpono i ka ʻenekinia kiko e like me nā cell seeder i loko o kahi ʻano o nā hoʻokolohua noiʻi kumu a me nā lāʻau lapaʻau.A hiki i kēia manawa, ʻaʻohe ʻano hana a me ke ʻano maʻamau no ka hoʻomalu maikaʻi ʻana o nā pūnaewele mesenchymal stem.Hiki i ka Countstar Rigel ke nānā i ka manaʻo, viability, a me nā hiʻohiʻona phenotype (a me kā lākou mau hoʻololi) i ka wā o ka hana ʻana a me ka hoʻokaʻawale ʻana o kēia mau pūnana pū.Loaʻa i ka Countstar Rigel ka pōmaikaʻi i ka loaʻa ʻana o ka ʻike morphological hou, i hāʻawi ʻia e ka brightfield mau a me nā hoʻopaʻa kiʻi fluorescence i ka wā o ke kaʻina holoʻokoʻa o ka nānā ʻana i ka maikaʻi o ke kelepona.Hāʻawi ka Countstar Rigel i kahi ala wikiwiki, paʻakikī, a hilinaʻi hoʻi no ka mālama ʻana i ka maikaʻi o nā pūnana pū.
Nā mea a me nā ʻano hana:
ʻO Adipose-derived mesenchymal stem cell (AdMSCs) i makana ʻia e Professor Nianmin Qi, AO/PI staining solution (Shanghai RuiYu, CF002).Antibody: CD29, CD34, CD45, CD56, CD73, CD105, HLADR (BD Company).
Hoʻoulu ʻia nā AdMSC i kahi incubator humidified 37 ℃, 5% CO2.E ʻeli me ka trypsin ma mua o ka hoʻohana ʻana.
Ua hahai ʻia ke kaʻina hana hoʻopaʻa CD marker e like me ka manual of antibody.
ʻIke ʻia ka māka CD me Countstar Rigel:
1. Ua hana ʻia kahi kaʻina hana hōʻailona kala ma ka hoʻonohonoho ʻana i ke kahawai PE i ke kiʻi PE fluorescence.
2. Ua hoʻopaʻa ʻia nā māla 3 mai kēlā me kēia keʻena.
3. Ma hope o ka pau ʻana o ke kiʻi a me ka nānā mua ʻana, ua hoʻonohonoho ʻia ka paepae (puka log) no ka hoʻololi maikaʻi a maikaʻi ʻole e ka polokalamu FCS.
ʻO ka hoʻomalu maikaʻi ʻana o ka pūnaʻi pū
Hōʻike kēia kiʻi (Figure 1) i ke kaʻina hana o lāʻau lāʻau stem cell .
Kiʻi 1: ʻO ke kaʻina hana no ka lāʻau lāʻau stem cell
Nā hualoaʻa:
Ka hoʻoholo ʻana i ka noʻonoʻo, hiki ke ola, ke anawaena, a me ka hōʻuluʻulu ʻana o AdMSC.
Ua hoʻoholo ʻia ke ola o AdMSC e AO/PI, Ua hana ʻia kahi kaʻina hana noi ʻelua kala ma ka hoʻonohonoho ʻana i ke kahawai ʻōmaʻomaʻo a me ke kahawai ʻulaʻula i ke kiʻi AO a me PI fluorescence, me kahi māla mālamalama.Hōʻike ʻia nā kiʻi laʻana ma ke Kiʻi 2.
Kiʻi 2. Nā kiʻi o mua o ka lawe ʻana a ma hope o ka lawe ʻana i nā AdMSC.A. Mamua o ka lawe ana;hōʻike ʻia kahi kiʻi hōʻike.B. Ma hope o ka lawe ʻana;hōʻike ʻia kahi kiʻi hōʻike.
Ua hoʻololi nui ʻia ka hiki ʻana o nā AdMSC ma hope o ka lawe ʻana i ka wā i hoʻohālikelike ʻia me mua o ka lawe ʻana.ʻO ka viability ma mua o ka lawe ʻana he 92%, akā ua hoʻemi ʻia i 71% ma hope o ka lawe ʻana.Ua hōʻike ʻia ka hopena ma ke Kiʻi 3.
Kiʻi 3. Nā hopena ola o nā AdMSC (Ma mua o ka lawe ʻana a ma hope o ka lawe ʻana)
Ua hoʻoholo pū ʻia ke anawaena a me ka hōʻuluʻulu ʻana e Countstar Rigel.Ua hoʻololi nui ʻia ke anawaena o nā AdMSC ma hope o ka lawe ʻana i ka wā i hoʻohālikelike ʻia me mua o ka lawe ʻana.ʻO ke anawaena ma mua o ka lawe ʻana he 19µm, akā ua hoʻonui ʻia i 21µm ma hope o ka lawe ʻana.ʻO ka hōʻuluʻulu o mua o ka lawe ʻana he 20%, akā ua hoʻonui ʻia i 25% ma hope o ka lawe ʻana.Mai nā kiʻi i paʻi ʻia e Countstar Rigel, ua hoʻololi nui ʻia ka phenotype o AdMSC ma hope o ka lawe ʻana.Ua hōʻike ʻia nā hopena ma ka Figure 4.
Kiʻi 4: ʻO ke anawaena a me nā hopena hōʻuluʻulu.A: Ua hoʻololi nui ʻia nā kiʻi hōʻike o AdMSCs, ka phenotype o AdMSCs ma hope o ka lawe ʻana.B: ʻO ka hōʻuluʻulu ma mua o ka lawe ʻana he 20%, akā ua hoʻonui ʻia i 25% ma hope o ka lawe ʻana.C: ʻO ke anawaena ma mua o ka lawe ʻana he 19µm, akā ua hoʻonui ʻia i 21µm ma hope o ka lawe ʻana.
E hoʻoholo i ka immunophenotype o AdMSC e Countstar Rigel
Ua hoʻoholo ʻia ka immunophenotype o AdMSC e Countstar Rigel, ua hoʻokomo ʻia nā AdMSC me nā antibody like ʻole (CD29, CD34, CD45, CD56, CD73, CD105, HLA-DR).Ua hana ʻia kahi kaʻina hana noi kala-hōʻailona ma ka hoʻonohonoho ʻana i kahi kahawai ʻōmaʻomaʻo e kiʻi i ka fluorescence PE, me kahi māla mālamalama.Ua hoʻohana ʻia ka māhele kiʻi kiʻi mālamalama ma ke ʻano he mask e hoʻāʻo i ka hōʻailona PE fluorescence.Ua hōʻike ʻia nā hopena o CD105 (Figure 5).
Kiʻi 5: Ua hoʻoholo ʻia nā hopena CD105 o AdMSC e Countstar Rigel.A: Ka helu helu o ka pākēneka maikaʻi o CD105 i nā laʻana like ʻole e FCS express 5 me ka lako polokalamu.B: Hāʻawi nā kiʻi kiʻekiʻe i ka ʻike morphological hou.C: Nā hualoaʻa i hōʻoia ʻia e nā liʻiliʻi liʻiliʻi o kēlā me kēia kelepona hoʻokahi, ua hoʻokaʻawale nā mea lako polokalamu FCS i nā cell i loko o nā pūʻulu like ʻole e like me kā lākou ʻano ʻano protein.
Ua hōʻike ʻia nā hopena antibodies ʻē aʻe ma Fig 6
Kiʻi 6: A: He kiʻi hōʻikeʻike o nā ASC me nā ʻano ʻano ʻano spindle maʻamau.Hopu ʻia e ka microscope OLYMPUS.Hoʻonui kumu, (10x).B: Hōʻike ʻia ka ʻokoʻa adipogenic o nā ASC e ka Ruthenium Red staining e hōʻike ana i nā wahi o ka mineralization.Hopu ʻia e ka microscope OLYMPUS.Hoʻonui kumu (10x).C: Countstar FL hiʻona o nā ASC.
Hōʻuluʻulu manaʻo:
Hiki i ka Countstar FL ke nānā i ka noʻonoʻo, hiki ke ola, a me nā hiʻohiʻona phenotype (a me kā lākou hoʻololi) i ka wā o ka hana ʻana a me ka hoʻokaʻawale ʻana o kēia mau pūnana pū.Hāʻawi ʻo FCS express i ka hana e nānā i kēlā me kēia cell signal, hōʻoia i ka ʻikepili ma o ke kiʻi.Hiki i ka mea hoʻohana ke hilinaʻi e hoʻokō i nā hoʻokolohua hou e pili ana i nā hopena Countstar Rigel.Hāʻawi ka Countstar Rigel i kahi ala wikiwiki, paʻakikī, a hilinaʻi hoʻi no ka mālama ʻana i ka maikaʻi o nā pūnana pū.
