Abstract: Mesenchymal stem masero inhengo yepluripotent stem maseru anogona kuparadzaniswa kubva kumesoderm.Nekuzvidzokorora kwavo patsva uye maitiro akasiyana-siyana ekusiyanisa, vane mukana wepamusoro wekurapa kwakasiyana-siyana mukurapa.Mesenchymal stem masero ane yakasarudzika immune phenotype uye immune regulation kugona.Naizvozvo, mesenchymal stem masero atove kushandiswa zvakanyanya mu stem cell transplantation, huinjiniya hwematishu, uye kuisirwa nhengo.Uye Kupfuura izvi maapplication, anoshandiswa sechishandiso chakakodzera muinjiniya yetishu seseder cell munhevedzano yezvidzidzo zvekutanga uye zvekiriniki.Kusvika parizvino, hapana nzira inogamuchirwa nevakawanda uye mwero wekutonga kwemhando yemesenchymal stem masero.Iyo Countstar Rigel inogona kutarisa kutarisisa, kugona, uye phenotype maitiro (uye shanduko dzadzo) panguva yekugadzira uye kusiyanisa kweaya stem maseru.Iyo Countstar Rigel ine zvakare mukana wekuwana yekuwedzera morphological ruzivo, yakapihwa neyakajeka nhandare uye fluorescence-yakavakirwa mifananidzo kurekodha panguva yese yekuongororwa kwemhando yesero.Iyo Countstar Rigel inopa inokurumidza, yakaomesesa, uye yakavimbika nzira yemhando yekutonga kwe stem maseru.
Zvishandiso uye nzira:
Adipose-yakatorwa mesenchymal stem masero (AdMSCs) vakapihwa zvipo naProfessor Nianmin Qi, AO/PI staining solution (Shanghai RuiYu, CF002).Antibody: CD29, CD34, CD45, CD56, CD73, CD105, HLADR (BD Company).
AdMSCs dzakagadzirwa mu37 ℃, 5% CO2 humidified incubator.Digest with trypsin usati washandisa.
CD marker staining process yakateverwa sebhuku rekudzivirira masoja.
CD marker yekuona ne Countstar Rigel:
1. Mucherechedzo-mavara ekushandisa maitiro akagadzirwa nekuisa PE chiteshi kumufananidzo PE fluorescence.
2. Minda mitatu yakatorwa kubva muimba imwe neimwe.
3. Mushure mekufungidzira uye kuongorora kwekutanga kwakapera, chikumbaridzo (log gedhi) chigadziro chekufambisa kwakanaka uye kwakashata kwakagadzirirwa neFCS software.
Kudzora kwemhando ye stem cell
Iyi inotevera Mufananidzo (Mufananidzo 1) inoratidza nzira ye stem cell therapy .
Mufananidzo 1: Maitiro e stem cell therapy
Mhinduro:
Kuona kutariswa, kugona, dhayamita, uye kuunganidzwa kweAdMSCs.
Kuvepo kweAdMSCs kwakatemwa neAO/PI, Maitiro ekunyorera-mavara maviri akagadzirwa nekuseta Green chiteshi uye Red chiteshi kumufananidzo AO nePI fluorescence, pamwe nemunda wakajeka.Mienzaniso yemifananidzo yakaratidzwa mumufananidzo 2.
Mufananidzo 2. Mifananidzo yepamberi yekutakura uye mushure mekutakura kweAdMSCs.A. Asati atakura;mufananidzo unomiririra unoratidzwa.B. Mushure mokufambisa;mufananidzo unomiririra unoratidzwa.
Iko kugona kweAdMSCs kwakashandurwa zvakanyanya mushure mekufambisa kana ichienzaniswa nepamberi pekutakura.Iko kugona pamberi pekutakura yaive 92%, asi yakadzikira kusvika 71% mushure mekufambisa.Mhedzisiro yakaratidzwa muFigure 3.
Mufananidzo 3. Mhedzisiro yekubudirira kweAdMSCs (Isati yatakura uye mushure mekufambisa)
Iyo dhayamita uye kuunganidzwa kwakatemwa zvakare na Countstar Rigel.Iyo dhayamita yeAdMSCs yakashandurwa zvakanyanya mushure mekufambisa kana ichienzaniswa nepamberi pekutakura.Iyo dhayamita isati yatakurwa yaive 19µm, asi yakawedzera kusvika 21µm mushure mekufambisa.Iko kuunganidzwa kwepamberi pekutakura yaive 20%, asi yakawedzera kusvika 25% mushure mekufambisa.Kubva pamifananidzo yakatorwa na Countstar Rigel, iyo phenotype yeAdMSCs yakashandurwa zvakanyanya mushure mekufambisa.Zvigumisiro zvakaratidzwa mumufananidzo 4.
Mufananidzo 4: The dhayamita uye aggregation zvabuda.A: Mifananidzo inomiririra yeAdMSCs, iyo phenotype yeAdMSCs yakashandurwa zvakanyanya mushure mekufambisa.B: Kuunganidzwa kusati kwatakura kwaive 20%, asi yakawedzera kusvika 25% mushure mekufambisa.C: Iyo dhayamita isati yatakurwa yaive 19µm, asi yakawedzera kusvika 21µm mushure mekufambisa.
Sarudza iyo immunophenotype yeAdMSCs naCountstar Rigel
Iyo immunophenotype yeAdMSCs yakatemerwa na Countstar Rigel, AdMSCs akaiswa maantibody akasiyana zvakateerana (CD29, CD34, CD45, CD56, CD73, CD105, HLA-DR).Iyo sign-color application maitiro akagadzirwa nekuisa Green chiteshi kumufananidzo PE fluorescence, pamwe nemunda wakajeka.Bright field reference segmentation yakashandiswa semask kuenzanisa chiratidzo chePE fluorescence.Migumisiro yeCD105 yakaratidzwa (Mufananidzo 5).
Mufananidzo 5: CD105 mhedzisiro yeAdMSCs yakatemwa naCountstar Rigel.A: Kuongororwa kwehuwandu hwechikamu chakanaka cheCD105 mumasampuli akasiyana neFCS express 5 plus software.B: Mifananidzo yemhando yepamusoro inopa mamwe morphological ruzivo.C: Mhedzisiro yakasimbiswa nezvigunwe zvesero rimwe nerimwe, maturusi esoftware eFCS akakamura maseru kuita mapoka akasiyana zvichienderana nekusiyana kwawo kweprotein expression.
Mamwe ma antibodies akaratidzwa muFig 6
Mufananidzo 6: A: Mufananidzo unomiririra weASCs ine yakajairwa spindle-shaped morphology.Yakatorwa neOLYMPUS microscope.Kukudza kwekutanga, (10x).B: Adipogenic kusiyanisa kweASCs kunoratidzwa neRuthenium Red staining inoratidza nzvimbo dze mineralization.Yakatorwa neOLYMPUS microscope.Kukudza kwekutanga (10x).C: Countstar FL maitiro eASCs.
Pfupiso:
Iyo Countstar FL inogona kutarisa kutarisisa, kugona, uye phenotype maitiro (uye shanduko dzadzo) panguva yekugadzira uye kusiyanisa kweaya stem maseru.FCS Express inopa basa rekuongorora sero rega rega, simbisa iyo data kuburikidza nemufananidzo.Mushandisi anogona zvakare kuve neruvimbo rwekuita zvinotevera zviedzo zvichibva pane Countstar Rigel zvawanikwa.Iyo Countstar Rigel inopa inokurumidza, yakaomesesa, uye yakavimbika nzira yemhando yekutonga kwe stem maseru.
