1.Yintoni inkcubeko yeseli?
Ukukhuliswa kweeseli kubhekiselele ekususeni iiseli kwizilwanyana okanye kwizityalo kwaye emva koko uzikhulisele kwindawo efanelekileyo eyenziweyo.Iiseli zinokuthathwa ngokuthe ngqo kwi-tissue kwaye ziphulwe ngeendlela ze-enzymatic okanye zoomatshini ngaphambi kokulima, okanye zinokuthi zithathwe kwimizila yeeseli ezimisiweyo okanye imigca yeeseli.
2.Yintoni inkcubeko yokuqala?
Inkcubeko esisiseko ibhekisa kwinqanaba lenkcubeko emva kokuba iiseli zihlulwe kwizicubu kwaye zande phantsi kweemeko ezifanelekileyo zide zithathe zonke ii-substrates ezikhoyo (oko kukuthi, ukufikelela kwi-confluence).Kweli nqanaba, iiseli kufuneka zikhuliswe kancinci ngokuzitshintshela kwisikhongozeli esitsha esinendawo entsha yokukhula ukubonelela ngesithuba esongezelelweyo sokukhula okuqhubekayo.
2.1 Umgca weselula
Emva kwe-subculture yokuqala, inkcubeko yokuqala ibizwa ngokuba yi-cell line okanye i-subclone.Imigca yeeseli ephuma kwiinkcubeko eziphambili zinobomi obulinganiselweyo (oko kukuthi zilinganiselwe; bona ngezantsi), kwaye njengoko zidlula, iiseli ezinomthamo ophezulu wokukhula zilawula, okukhokelela kwinqanaba elithile le-genotype kuluntu olungagungqiyo kwi-phenotype.
2.2 Uxinzelelo lweeseli
Ukuba i-subpopulation yomgca weseli ikhethwe ngokufanelekileyo kwinkcubeko ngokwenza i-cloning okanye enye indlela, umgca weseli uya kuba luxinzelelo lweseli.Iintlobo zeeseli zihlala zifumana utshintsho olongezelelweyo lwemfuzo emva kokuba umgca wabazali uqalile.
3.Imigca yeseli enyiniweyo kwaye eqhubekayo
Iiseli eziqhelekileyo ngokuqhelekileyo zahlula inani elilinganiselweyo lamaxesha ngaphambi kokuba ziphulukane namandla okwandisa.Esi sisiganeko esimiselwe ngokwemfuza esibizwa ngokuba yi-senescence;le migca yeeseli ibizwa ngokuba yimigca yeeseli egqityiweyo.Nangona kunjalo, ezinye iiseli zeeseli ziyafa ngenxa yenkqubo ebizwa ngokuba yinguqu, enokuthi yenzeke ngokuzenzekelayo okanye enokuthi yenziwe yimichiza okanye iintsholongwane.Xa umgca weseli ogqityiweyo wenza utshintsho kwaye ufumana isakhono sokwahlula ngokungenammiselo, uba ngumgca weseli oqhubekayo.
4.Imeko yenkcubeko
Iimeko zenkcubeko zohlobo lweseli nganye zahluke kakhulu, kodwa indawo eyenziweyo yeeseli zokulima isoloko iqulunqwe sisitya esifanelekileyo, esiqulethe oku kulandelayo:
4.1 I-Substrate okanye i-culture medium ebonelela ngezondlo eziyimfuneko (i-amino acids, iicarbohydrates, iivithamini, iiminerali)
4.2 Imiba yokukhula
4.3 IiHormones
4.4 Iigesi (O2, CO2)
4.5 Ukulawulwa kokusingqongileyo ngokomzimba kunye neekhemikhali (pH, uxinzelelo lwe-osmotic, ubushushu)
Uninzi lweeseli zixhomekeke kwi-anchorage kwaye kufuneka zikhuliswe kwi-substrate eqinileyo okanye i-semi-solid substrate (inkcubeko yokubambelela okanye i-monolayer), ngelixa ezinye iiseli zinokukhula zidada phakathi (inkcubeko yokunqunyanyiswa).
5.Cryopreservation
Ukuba kukho iiseli ezigqithisileyo kwi-subculture, kufuneka ziphathwe nge-agent ekhuselekileyo yokukhusela (njenge-DMSO okanye i-glycerol) kwaye igcinwe kwiqondo lokushisa elingaphantsi kwe-130 ° C (cryopreservation) de zifuneka.Ngolwazi oluthe kratya malunga ne-subculture kunye ne-cryopreservation yeeseli.
6.I-morphology yeeseli kwinkcubeko
Iiseli ngokwenkcubeko zinokwahlulwa zibe ziindidi ezintathu ezisisiseko ngokwemilo kunye nenkangeleko yazo (okt i-morphology).
6.1 Iiseli ze-Fibroblasts zine-bipolar okanye i-multipolar, zinemilo emide, kwaye zikhula ziqhotyoshelwe kwi-substrate.
6.2 Iiseli ezinjenge-epithelial zi-polygonal, zinesayizi eqhelekileyo, kwaye zifakwe kwi-matrix kumaphepha adiscrete.
6.3 Iiseli ezinjenge-Lymphoblast zingqukuva kwaye zihlala zikhula ngokunqunyanyiswa ngaphandle kokuncamathela kumphezulu.
7.Ukusetyenziswa kwenkcubeko yeeseli
Ukukhuliswa kweeseli sesinye sezixhobo eziphambili ezisetyenziswa kwiiseli kunye nebhayoloji yemolekyuli.Ibonelela ngenkqubo yemodeli egqwesileyo yokufunda i-physiology eqhelekileyo kunye ne-biochemistry yeeseli (ezifana nophando lwe-metabolic, ukuguga), iimpembelelo zamachiza kunye ne-toxic compounds kwiiseli, kunye ne-mutagenesis kunye ne-carcinogenic effects.Ikwasetyenziselwa ukukhangela amachiza kunye nophuhliso kunye nokuveliswa okukhulu kweekhompawundi zebhayoloji (ezifana nezitofu zokugonya, iiproteni zonyango).Inzuzo ephambili yokusebenzisa inkcubeko yeseli kuyo nayiphi na yezi zicelo kukuhambelana kunye nokuveliswa kwakhona kweziphumo ezinokufumaneka ngokusebenzisa ibhetshi yeeseli ezidibeneyo.
Ixesha lokuposa: Jun-03-2019