Nkọwa: Mesenchymal stem cell bụ akụkụ nke mkpụrụ ndụ stem pluripotent nke enwere ike kewapụ na mesoderm.Site na mmeghari ohuru nke onwe ha na njirimara di iche iche nke uzo, ha nwere ikike di elu maka usoro ogwu di iche iche na ogwu.Selụ sel mesenchymal nwere phenotype nke na-alụso ọrịa ọgụ na ikike ịchịkwa usoro ahụ ji alụso ọrịa ọgụ.Ya mere, a na-eji mkpụrụ ndụ mesenchymal eme ihe n'ọtụtụ ebe na ntụgharị cell cell transplantation, nhazi anụ ahụ, na ntụgharị akụkụ ahụ.Na gafere ngwa ndị a, a na-eji ha dị ka ezigbo ngwá ọrụ na injinịa anụ ahụ dị ka mkpụrụ ndụ mkpụrụ osisi na usoro nyocha nyocha nke isi na nke ụlọ ọgwụ.Ruo ugbu a, ọ nweghị usoro na ụkpụrụ a na-anabata nke ọma maka njikwa mma nke sel sel mesenchymal.Countstar Rigel nwere ike nyochaa itinye uche, ike, na njirimara phenotype (na mgbanwe ha) n'oge mmepụta na ọdịiche nke sel ndị a.Countstar Rigel nwekwara uru dị n'inweta ozi morphological ndị ọzọ, nke na-enye site na ndekọ onyonyo na-adịgide adịgide na nke dabere na fluorescence n'oge usoro nleba anya ogo cell.Countstar Rigel na-enye usoro ngwa ngwa, ọkaibe na nke a pụrụ ịdabere na ya maka njikwa mma nke sel sel.
Akụrụngwa na ụzọ:
Mkpụrụ ndụ mesenchymal mesenchymal adipose (AdMSCs) nyere onyinye site n'aka Prọfesọ Nianmin Qi, ngwọta ntụpọ AO/PI (Shanghai RuiYu, CF002).Ihe mgbochi: CD29, CD34, CD45, CD56, CD73, CD105, HLADR (Ụlọ ọrụ BD).
Azụlitere AdMSC na 37℃, 5% CO2 incubator humidified.Gwakọta ya na trypsin tupu ejiri ya.
A na-agbaso usoro ntụpọ CD dị ka akwụkwọ ntuziaka nke mgbochi.
Nchọpụta akara CD na Countstar Rigel:
1. Emepụtara usoro ntinye akara ngosi site na ịtọ ọwa PE ka ọ bụrụ ihe onyonyo PE fluorescence.
2. E weghaara ubi 3 n'ime ụlọ nke ọ bụla.
3. Mgbe nyochachara onyonyo na nyocha mbụ, ntọala ọnụ ụzọ (ọnụ ụzọ log) maka nfefe dị mma na nke na-adịghị mma ka edobere ngwa ngwa FCS.
Njikwa ogo nke cell stem
Ọnụ ọgụgụ na-esonụ (Fig 1) na-egosi usoro nke ọgwụgwọ sel stem .
Ọgụgụ 1: Usoro maka ọgwụgwọ sel stem
Nsonaazụ:
Na-ekpebi itinye uche, ịdị ndụ, dayameta, na nchịkọta nke AdMSC.
AO/PI kpebisiri ike ịdị adị nke AdMSC, emebere usoro ngwa nwere agba abụọ site na ịtọ chawa Green na ọwa uhie ka ọ bụrụ onyonyo AO na PI fluorescence, gbakwunyere ubi na-egbuke egbuke.E gosiri onyonyo atụ na eserese 2.
Onyonyo 2. Foto nke tupu ebuga na mgbe njem nke AdMSC gasịrị.A. Tupu ebuga;egosiri ihe onyonyo nnochite anya.B. Mgbe ụgbọ njem gasịrị;egosiri ihe onyonyo nnochite anya.
A gbanwere ike nke AdMSC nke ukwuu mgbe ụgbọ njem gachara ma atụnyere tupu ebufee ya.The viability tupu njem bụ 92%, ma ọ belatara 71% mgbe ụgbọ njem.E gosipụtara nsonaazụ ya na eserese 3.
Ọgụgụ 3. Nsonaazụ viability nke AdMSCs (Tupu ebufe na mgbe ụgbọ njem gasịrị)
Countstar Rigel kpebikwara dayameta na mkpokọta.Agbanwere dayameta nke AdMSC nke ukwuu mgbe ụgbọ njem gachara ma atụnyere tupu ebufee ya.Dayameta tupu ebuga ya bụ 19µm, mana ọ gbagoro na 21µm mgbe ụgbọ njem gachara.Nchịkọta nke tupu ebuga bụ 20%, mana ọ mụbara ruo 25% mgbe ụgbọ njem gasịrị.Site na onyonyo Countstar Rigel weghaara, phenotype nke AdMSC gbanwere nke ukwuu mgbe ụgbọ njem gachara.E gosipụtara nsonaazụ ya na eserese 4.
Ọnụọgụ 4: The dayameta na nchịkọta nchịkọta.A: Onyonyo nnọchiteanya nke AdMSC, phenotype nke AdMSC gbanwere nke ukwuu mgbe ụgbọ njem gasịrị.B: Nchịkọta tupu ebuga bụ 20%, mana ọ mụbara ruo 25% mgbe ụgbọ njem gasịrị.C: Dayameta tupu ebufe ya bụ 19µm, mana ọ gbagoro na 21µm mgbe ụgbọ njem gachara.
Kpebie ụdị immunophenotype nke AdMSC site n'aka Countstar Rigel
Countstar Rigel kpebisiri ike na ụdị immunophenotype nke AdMSC, ndị AdMSC bu ụzọ mgbochi dị iche iche (CD29, CD34, CD45, CD56, CD73, CD105, HLA-DR).Emepụtara usoro ngwa mgbaama site na ịtọ ọwa Green ka ọ nwee onyonyo PE fluorescence, gbakwunyere ubi na-egbuke egbuke.A na-etinye akụkụ ntụaka ihe onyonyo na-egbuke egbuke dị ka ihe nkpuchi iji gosipụta akara ngosi PE fluorescence.E gosiputara nsonaazụ CD105 (Foto 5).
Ọgụgụ 5: Countstar Rigel kpebiri nsonaazụ CD105 nke AdMSC.A: Ntụle ọnụ ọgụgụ dị mma nke CD105 na nlele dị iche iche site na FCS express 5 plus software.B: Onyonyo dị elu na-enye ozi morphological ọzọ.C: Nsonaazụ enwetara site na mkpịsị mkpịsị mkpịsị aka nke otu cell ọ bụla, ngwa ngwa FCS kewara sel ndị ahụ n'ime otu dị iche iche dịka okwu protein ha dị iche iche siri dị.
E gosipụtara nsonaazụ ọgwụ mgbochi ndị ọzọ na Fig 6
Onyonyo 6: A: Onyonyo nnọchite anya ASC nwere ụdị njiri mara ụdị spindle.Igwe microscope OLYMPUS weghaara ya.Mmalite izizi, (10x).B: A na-egosipụta ọdịiche Adipogenic nke ASC site na Ruthenium Red staining na-egosi ebe ịnweta mineral.Igwe microscope OLYMPUS weghaara ya.Mmalite izizi (10x).C: Nkọwapụta Countstar FL nke ASC.
Nchịkọta:
Countstar FL nwere ike nyochaa nleba anya, ịdị ndụ, na njirimara phenotype (na mgbanwe ha) n'oge mmepụta na ọdịiche nke sel ndị a.FCS express na-enye ọrụ iji nyochaa cell mgbaama ọ bụla, kwado data ahụ site na onyonyo a.Onye ọrụ nwekwara ike inwe obi ike ime nnwale ndị ọzọ dabere na nsonaazụ Countstar Rigel.Countstar Rigel na-enye usoro ngwa ngwa, ọkaibe na nke a pụrụ ịdabere na ya maka njikwa mma nke sel sel.
