Imwe yemabhenefiti makuru ehunhu hwesero kugona kushandura kemikari yemuviri yekuberekana kwesero (kureva tembiricha, pH, osmotic pressure, O2 uye CO2 tension) uye physiological environment (kureva mahormone uye kudya kunovaka muviri).Mukuwedzera kune tembiricha, nharaunda yetsika inodzorwa nekukura kwepakati.
Kunyangwe iyo physiological nharaunda yetsika isina kujeka seyayo yemuviri uye kemikari nharaunda, kunzwisiswa kuri nani kweserum zvikamu, kuzivikanwa kwekukura kwezvinhu zvinodiwa pakuwedzera, uye kunzwisisa kuri nani kweiyo microenvironment yemasero mutsika.(Kureva kupindirana kwesero-sero, kufambiswa kwegasi, kudyidzana nematrix) ikozvino inobvumira mamwe maseru mitsara kuti agadziriswe muserum-isina midhiya.
1.Culture nharaunda inokanganisa sero kukura
Ndokumbira utarise kuti sero tsika mamiriro akasiyana kune yega sero mhando.
Mibairo yekutsauka kubva kumagariro mamiriro anodiwa kune rumwe rudzi rwesero kubva pakutaura kweabnormal phenotypes kusvika kukundikana zvachose kwesero tsika.Naizvozvo, isu tinokurudzira kuti iwe ujairane nesero mutsara wauri kufarira uye kunyatsotevera mirairo inopihwa kune yega chigadzirwa chaunoshandisa mukuyedza kwako.
2.Precautions yekugadzira yakagadziridzwa sero tsika nharaunda yemasero ako:
Tsika midhiya uye serum (ona pazasi kuti uwane rumwe ruzivo)
pH uye CO2 mazinga (ona pazasi kuti uwane rumwe ruzivo)
Rima mapurasitiki (ona pazasi kuti uwane rumwe ruzivo)
Tembiricha (ona pazasi kuti uwane rumwe ruzivo)
2.1 Cultural Media uye Serum
Iyo tsika yepakati ndiyo inonyanya kukosha yenzvimbo yetsika, nekuti inopa chikafu, kukura zvinhu uye mahomoni anodiwa pakukura kwesero, uye inogadzirisa iyo pH uye osmotic pressure yetsika.
Kunyangwe iyo yekutanga sero tsika kuyedza yakaitwa pachishandiswa echisikigo midhiya inotorwa kubva munyama inotorwa uye mvura yemuviri, kudiwa kwekumisikidzwa, mhando yenhau, uye kuwedzera kudiwa kwakatungamira mukuvandudzwa kweiyo midhiya.Iwo matatu marudzi ekutanga enhau ndeye basal media, yakaderedzwa serum midhiya uye serum-yemahara midhiya, uye vane zvakasiyana zvinodiwa zvekuwedzera serum.
2.1.1 Basic medium
Gibco cell tsika yepakati
Mazhinji maseru mitsara anokura zvakanaka mumidhiya yekutanga ine amino acids, mavhitaminzi, inorganic munyu, uye kabhoni masosi (akadai seglucose), asi aya ekutanga midhiya magadzirirwo anofanirwa kuwedzerwa neserum.
2.1.2 Yakaderedzwa serum medium
Bhodhoro neGibco Low Serum Medium
Imwe nzira yekudzikisa yakashata mhedzisiro ye serum mune yesero tsika kuyedza ndeye kushandisa serum-yakaderedzwa midhiya.Yakaderedzwa serum yepakati ndiyo yakakosha yepakati formula yakapfuma mune zvinovaka muviri uye zvinhu zvinobva kumhuka, izvo zvinogona kuderedza huwandu hweserum inodiwa.
2.1.3 Serum-isina svikiro
Bhodhoro neGibco serum-yemahara yepakati
Serum-isina svikiro (SFM) inotenderedza kushandiswa kweserum yemhuka nekutsiva serum nekudya kwakakodzera uye kuumbwa kwehomoni.Tsika dzakawanda dzekutanga uye maseru mitsara ine serum-yemahara yepakati magadzirirwo, anosanganisira Chinese Hamster Ovary (CHO) recombinant protein yekugadzira mutsara, akasiyana hybridoma cell mitsara, mitsara yezvipembenene Sf9 uye Sf21 (Spodoptera frugiperda), pamwe neyeMugadziri wekugadzira hutachiona. (somuenzaniso, 293, VERO, MDCK, MDBK), nezvimwewo. Chimwe chezvakakosha zvekushandisa serum-isina svikiro ndiko kukwanisa kuita svikiro kusarudzwa kune mamwe marudzi emasero nekusarudza kusanganiswa kwakakodzera kwekukura kwezvinhu.Tafura inotevera inodonongodza zvakanakira uye zvakaipira zveserum-yemahara media.
Advantage
Wedzera kujeka
Kuwedzera kuenderana kuita
Kucheneswa kuri nyore uye kudzika kwemvura kugadzirisa
Kunyatsoongorora kushanda kwesero
Wedzera kubereka
Kudzora kurinani kwemaitiro emuviri
Yakanatsiridza nharembozha kuona
Kukanganisa
Cell type chaiyo yepakati formula inodiwa
Inoda yakakwirira reagent kuchena
Kuderera mukukura
2.2.1 pH level
Mazhinji akajairwa mammalian cell mitsara inokura zvakanaka papH 7.4, uye mutsauko uripo pakati pemitsara yesero idiki.Zvisinei, mamwe mitsara yesero yakashandurwa yakaratidzwa kukura zviri nani munzvimbo ine acidic zvishoma (pH 7.0 - 7.4), nepo mamwe mafibroblast sero mitsara akajairwa anofarira zvishoma alkaline nharaunda (pH 7.4 - 7.7).Insect cell mitsara seSf9 neSf21 inokura zvakanyanya papH 6.2.
2.2.2 CO2 nhanho
Iyo yekukura yepakati inodzora pH yetsika uye inovharira maseru mutsika kuramba shanduko mupH.Kazhinji, kubhafa uku kunowanikwa neine organic (semuenzaniso, HEPES) kana CO2-bicarbonate-based buffers.Nekuda kwekuti pH yepakati inotsamira pachiyero chakapfava chekabhoni dhaikisaidhi (CO2) uye bicarbonate (HCO3-), shanduko mumhepo CO2 inoshandura pH yepakati.Naizvozvo, kana uchishandisa svikiro yakavharwa ne CO2-bicarbonate-based buffer, zvinofanirwa kushandisa exogenous CO2, kunyanya kana culturing masero muakavhurika tsika ndiro kana culturing yakashandurwa sero mitsetse pakuwanda kwakanyanya.Kunyange zvazvo vatsvakurudzi vakawanda vanowanzoshandisa 5-7% CO2 mumhepo, miedzo yakawanda yemasero inowanzoshandisa 4-10% CO2.Zvisinei, svikiro imwe neimwe ine CO2 tension inokurudzirwa uye bicarbonate concentration kuti iwane pH yakarurama uye osmotic pressure;kuti uwane rumwe ruzivo, ndapota tarisa kune mirairo yemugadziri wepakati.
2.3 Kurima mapurasitiki
Cell tsika mapurasitiki anowanikwa mune akasiyana mafomu, saizi uye nepamusoro kuti aenderane akasiyana siyana sero tsika yekushandisa.Shandisa sero tsika epurasitiki pamusoro pegwara uye sero tsika chigaba chegwara pazasi kuti ikubatsire kusarudza iyo purasitiki yakakodzera kune yako cell tsika application.
Wona ese Thermo Scientific Nunc cell tsika mapurasitiki (advertising link)
2.4 Tembiricha
Iyo yakanyanya tembiricha yetsika yesero inoenderana zvakanyanya nekupisa kwemuviri wemugamuchiri kubva kune iyo masero ari ega, uye zvishoma pane yeanatomical shanduko yekushisa (semuenzaniso, tembiricha yeganda inogona kunge yakaderera pane yeskeletal tsandanyama. )Kune sero tsika, Kupisa idambudziko rakakura kupfuura kupisa.Naizvozvo, tembiricha iri muincubator inowanzoiswa zvishoma pazasi peyakanyanya tembiricha.
2.4.1 Optimum tembiricha dzakasiyana sero mitsetse
Mitsetse yakawanda yevanhu neinoyamwisa inochengetwa pa36°C kusvika 37°C kuti ikure zvakanaka.
Masero ezvipembenene anorimwa pa27 ° C kuti awedzere kukura;dzinokura zvishoma nezvishoma pane tembiricha dzakaderera uye tembiricha dziri pakati pe27°C ne30°C.Pamusoro pe30 ° C, simba remasero ezvipembenene rinoderera, kunyange kana richidzokera ku27 ° C, masero haazodzoki.
Avian cell line inoda 38.5°C kuti isvike pakukura.Kunyange zvazvo masero aya achigona kuchengetwa pa37°C, anokura zvishoma nezvishoma.
Mitsetse yemasero anobva kumhuka dzine ropa rinotonhora (semhuka dzemufibhi, hove dzemvura inotonhora) dzinogona kushivirira tembiricha yakawanda ye15°C kusvika 26°C.
Nguva yekutumira: Feb-01-2023