1.Iyini isiko lamaseli?
I-cell culture isho ukukhipha amaseli ezilwaneni noma ezitshalweni bese uwakhulisa endaweni yokwenziwa evumayo.Amaseli angathathwa ngokuqondile esicutshini futhi ahlakazwe ngendlela ye-enzymatic noma yemishini ngaphambi kokulima, noma angathathwa emigqeni yamaseli emisiwe noma imigqa yamaseli.
2.Yini isiko eliyisisekelo?
Isiko eliyisisekelo lisho isigaba sempucuko ngemva kokuba amaseli ehlukene nezicubu futhi anda ngaphansi kwezimo ezifanele aze athathe wonke ama-substrates atholakalayo (okungukuthi, ukufinyelela ukuhlangana).Kulesi sigaba, amangqamuzana kufanele enziwe kancane ngokuwadlulisela esitsheni esisha esinendawo yokukhula entsha ukuze anikeze isikhala esengeziwe sokukhula okuqhubekayo.
2.1 Ulayini weselula
Ngemuva kwe-subculture yokuqala, isiko eliyisisekelo libizwa ngokuthi i-cell line noma i-subclone.Imigqa yamaseli etholwe kumasiko ayisisekelo anesikhathi esilinganiselwe sokuphila (okungukuthi anomkhawulo; bheka ngezansi), futhi njengoba edlula, amaseli anamandla okukhula aphezulu ayabusa, okuholela ezingeni elithile le-genotype kubantu abaphikisana ne-phenotype.
2.2 Ubunzima beseli
Uma ukugcwala okuncane komugqa weseli kukhethwa kahle kusiko ngokwenza i-cloning noma enye indlela, ulayini weseli uzoba uhlobo lweseli.Izinhlobo zamaseli ngokuvamile zithola izinguquko ezengeziwe zofuzo ngemva kokuqala komugqa wabazali.
3.Olayini beselula abanomkhawulo futhi abaqhubekayo
Amaseli avamile ngokuvamile ahlukanisa inani elilinganiselwe lezikhathi ngaphambi kokulahlekelwa amandla okwanda.Lesi isenzakalo esinqunywa ngofuzo esibizwa ngokuthi i-senescence;le migqa yamaseli ibizwa ngokuthi imigqa yamaseli alinganiselwe.Kodwa-ke, amanye amangqamuzana agcina engafi ngenqubo ebizwa ngokuthi ukuguqulwa, engenzeka ngokuzenzakalelayo noma engenziwa amakhemikhali noma amagciwane.Uma umugqa weseli olinganiselwe ushintsha futhi uthola ikhono lokuhlukanisa unomphela, uba umugqa weseli oqhubekayo.
4.Isimo sesiko
Izimo zesiko zohlobo lweseli ngalinye zihluke kakhulu, kodwa indawo yokwenziwa yamaseli okulima ihlale inesitsha esifanelekile, esiqukethe lokhu okulandelayo:
4.1 I-substrate noma i-culture medium ehlinzeka ngemisoco ebalulekile (ama-amino acid, ama-carbohydrate, amavithamini, amaminerali)
4.2 Izici zokukhula
4.3 Amahomoni
4.4 Amagesi (O2, CO2)
4.5 Imvelo elawulwayo ngokomzimba namakhemikhali (pH, ingcindezi ye-osmotic, izinga lokushisa)
Amaseli amaningi ancike ku-anchorage futhi kufanele akhuliswe ku-substrate eqinile noma eqinile kancane (isiko lokunamathela noma le-monolayer), kuyilapho amanye amaseli angakhula entanta phakathi nendawo (isiko lokumiswa).
5.Cryopreservation
Uma kunamangqamuzana adlulele ku-subculture, kufanele aphathwe nge-ejenti yokuvikela efanelekile (njenge-DMSO noma i-glycerol) futhi agcinwe ezingeni lokushisa elingaphansi -130 ° C (cryopreservation) kuze kube yilapho edingeka.Ukuze uthole ulwazi olwengeziwe mayelana ne-subculture kanye ne-cryopreservation yamaseli.
6.I-morphology yamaseli esikweni
Amaseli esikweni angahlukaniswa abe izigaba ezintathu eziyisisekelo ngokusekelwe ekubunjweni kwawo kanye nokubukeka kwawo (okungukuthi i-morphology).
6.1 Amaseli e-Fibroblasts ane-bipolar noma ama-multipolar, anomumo obude, futhi akhula enamathele ku-substrate.
6.2 Amaseli afana ne-epithelial ane-polygonal, anosayizi ovamile, futhi anamathiselwe ku-matrix ngamashidi ahlukene.
6.3 Amaseli afana ne-Lymphoblast ayindilinga futhi ngokuvamile akhula ngokumiswa ngaphandle kokunamathela phezulu.
7.Ukusetshenziswa kwesiko lamaseli
Isiko lamaseli lingelinye lamathuluzi ayinhloko asetshenziswa kubhayoloji yamangqamuzana nengqamuzana.Ihlinzeka ngohlelo lwemodeli oluhle kakhulu lokufunda i-physiology evamile kanye ne-biochemistry yamaseli (njengocwaningo lwe-metabolic, ukuguga), imiphumela yezidakamizwa nezinhlanganisela ezinobuthi kumaseli, kanye nemiphumela ye-mutagenesis kanye ne-carcinogenic.Ibuye isetshenziselwe ukuhlola nokuthuthukiswa kwezidakamizwa kanye nokukhiqizwa okukhulu kwezinhlanganisela zezinto eziphilayo (njengemithi yokugoma, amaprotheni okwelapha).Inzuzo enkulu yokusebenzisa isiko lamaseli kunoma yisiphi salezi zinhlelo zokusebenza ukuvumelana kanye nokuphindaphindeka kwemiphumela engatholwa kusetshenziswa iqoqo lamaseli ahlanganisiwe.
Isikhathi sokuthumela: Jun-03-2019