Ɗaya daga cikin manyan fa'idodin al'adar tantanin halitta shine ikon sarrafa sinadarai na zahiri na haifuwar tantanin halitta (watau zazzabi, pH, matsa lamba osmotic, tashin hankali O2 da CO2) da yanayin physiological (watau hormone da maida hankali na gina jiki).Bugu da ƙari ga zafin jiki, yanayin al'ada yana sarrafawa ta hanyar haɓaka.
Ko da yake yanayin yanayin ilimin lissafi na al'ada ba shi da kyau kamar yanayin jiki da sinadarai, mafi kyawun fahimtar abubuwan da ke tattare da kwayoyin halitta, gano abubuwan haɓaka da ake buƙata don haɓakawa, da kuma fahimtar ƙananan ƙwayoyin sel a cikin al'ada.(Watau hulɗar tantanin halitta, yaduwar iskar gas, hulɗa tare da matrix) yanzu yana ba da damar haɓaka wasu layukan tantanin halitta a cikin kafofin watsa labarai marasa amfani.
1.Culture muhalli yana shafar ci gaban cell
Lura cewa yanayin al'adun tantanin halitta sun bambanta ga kowane nau'in tantanin halitta.
Sakamakon karkacewa daga yanayin al'ada da ake buƙata don wani nau'in tantanin halitta ya bambanta daga bayyanar da abubuwan da ba su dace ba zuwa ga cikakkiyar gazawar al'adun tantanin halitta.Don haka, muna ba da shawarar cewa ku saba da layin tantanin halitta da kuke sha'awar kuma ku bi ƙa'idodin da aka bayar don kowane samfurin da kuke amfani da shi a gwajin ku.
2.Trecautions don ƙirƙirar ingantaccen yanayin al'adun tantanin halitta don ƙwayoyin ku:
Kafofin watsa labarai na al'ada da magani (duba ƙasa don ƙarin bayani)
pH da CO2 matakan (duba ƙasa don ƙarin bayani)
Noma filastik (duba ƙasa don ƙarin bayani)
Zazzabi (duba ƙasa don ƙarin bayani)
2.1 Kafofin watsa labarai na Al'adu da Magani
Matsakaicin al'ada shine mafi mahimmancin yanayin yanayin al'ada, saboda yana ba da abinci mai gina jiki, abubuwan haɓakawa da hormones da ake buƙata don haɓakar ƙwayoyin cuta, kuma yana daidaita pH da matsa lamba na osmotic na al'ada.
Kodayake an gudanar da gwaje-gwajen al'adun cell na farko ta hanyar amfani da kafofin watsa labaru na halitta da aka samo daga kayan aikin nama da ruwan jiki, buƙatar daidaitawa, ingancin kafofin watsa labaru, da karuwar buƙatun ya haifar da ci gaban ingantaccen kafofin watsa labaru.Nau'o'in kafofin watsa labarai guda uku na asali sune kafofin watsa labarai na basal, rage yawan kafofin watsa labarai da kafofin watsa labarai marasa amfani, kuma suna da buƙatu daban-daban don ƙarin maganin jini.
2.1.1 Matsakaici na asali
Gibco cell al'ada matsakaici
Yawancin layukan tantanin halitta suna girma da kyau a cikin kafofin watsa labarai na asali waɗanda ke ɗauke da amino acid, bitamin, salts inorganic, da maɓuɓɓugar carbon (kamar glucose), amma dole ne a ƙara waɗannan ƙa'idodin kafofin watsa labarai da magani.
2.1.2 Rage matsakaicin magani
Kwalba tare da Gibco Low Serum Medium
Wata dabarar da za ta rage illar cutar sinadari a cikin gwaje-gwajen al'adun tantanin halitta ita ce yin amfani da kafofin watsa labarai da aka rage masu.Rage matsakaicin ƙwayar magani shine ainihin maƙasudin matsakaici mai wadata a cikin abubuwan gina jiki da abubuwan da aka samo daga dabba, wanda zai iya rage adadin maganin da ake buƙata.
2.1.3 Matsakaicin mara amfani da magani
Kwalba tare da Gibco matsakaici mara maganin magani
Matsakaicin marasa lafiya (SFM) yana kewaye da amfani da maganin dabba ta hanyar maye gurbin magani tare da ingantaccen abinci mai gina jiki da tsarin hormone.Yawancin al'adu na farko da layin tantanin halitta suna da matsakaicin matsakaici mara amfani, gami da layin samar da furotin na Hamster Ovary (CHO), layin ƙwayoyin ƙwayoyin cuta daban-daban, layin kwari Sf9 da Sf21 (Spodoptera frugiperda), da kuma mai watsa shiri don samar da ƙwayoyin cuta. (misali, 293, VERO, MDCK, MDBK), da dai sauransu. Ɗaya daga cikin manyan fa'idodin yin amfani da matsakaicin matsakaici mara amfani shine ikon yin matsakaicin zaɓi don takamaiman nau'ikan tantanin halitta ta hanyar zaɓar abubuwan da suka dace na abubuwan haɓaka.Tebur mai zuwa yana lissafin fa'idodi da rashin amfani na kafofin watsa labarai marasa magani.
Amfani
Ƙara haske
Ƙarin daidaiton aiki
Sauƙin tsarkakewa da sarrafa ƙasa
Auna aikin tantanin halitta daidai
Ƙara yawan aiki
Kyakkyawan kula da halayen ilimin lissafi
Ingantattun gano kafofin watsa labaru
Hasara
Sel irin takamaiman matsakaicin dabara bukatun
Bukatar mafi girma reagent tsarki
Ragewar girma
2.2.1 pH
Yawancin layukan tantanin halitta na dabbobi masu shayarwa suna girma da kyau a pH 7.4, kuma bambance-bambance tsakanin layin salula daban-daban kadan ne.Duk da haka, an nuna wasu layukan tantanin halitta da aka canza don girma mafi kyau a cikin dan kadan acidic yanayi (pH 7.0 - 7.4), yayin da wasu layukan fibroblast na yau da kullum sun fi son yanayin alkaline kadan (pH 7.4 - 7.7).Layukan ƙwayoyin kwari kamar Sf9 da Sf21 suna girma mafi kyau a pH 6.2.
2.2.2 CO2 matakin
Matsakaicin haɓaka yana sarrafa pH na al'ada kuma yana ɓoye sel a cikin al'ada don tsayayya da canje-canje a pH.Yawancin lokaci, ana samun wannan buffer ta hanyar ƙunsar kwayoyin halitta (misali, HEPES) ko tushen buffers na tushen CO2-bicarbonate.Saboda pH na matsakaici ya dogara da ma'auni mai laushi na narkar da carbon dioxide (CO2) da bicarbonate (HCO3-), canje-canje a cikin CO2 na yanayi zai canza pH na matsakaici.Sabili da haka, lokacin amfani da matsakaicin buffer tare da buffer na tushen CO2-bicarbonate, ya zama dole a yi amfani da CO2 exogenous, musamman lokacin da ake yin sel a cikin buɗaɗɗen abinci na al'ada ko al'adar da aka canza layin salula a babban taro.Kodayake yawancin masu bincike yawanci suna amfani da 5-7% CO2 a cikin iska, yawancin gwaje-gwajen al'adun tantanin halitta yawanci suna amfani da 4-10% CO2.Duk da haka, kowane matsakaici yana da shawarar CO2 tashin hankali da bicarbonate maida hankali don cimma daidai pH da osmotic matsa lamba;don ƙarin bayani, da fatan za a koma zuwa umarnin masana'anta na matsakaici.
2.3 Noman robobi
Ana samun robobin al'adar salula ta nau'i-nau'i, girma da sama don dacewa da aikace-aikacen al'adun tantanin halitta daban-daban.Yi amfani da jagorar shimfidar filaye na al'adar tantanin halitta da jagorar kwantenan al'adun tantanin halitta da ke ƙasa don taimaka muku zaɓin filastik da ya dace don aikace-aikacen al'adun cell ɗin ku.
Duba duk robobin al'adun cell na Thermo Scientific Nunc (hanyar talla)
2.4 Zazzabi
Mafi kyawun zafin jiki don al'adar tantanin halitta ya dogara da babban yanayin zafin jiki na mai watsa shiri wanda sel ke ware, kuma zuwa ƙaramin adadin canje-canjen yanayin zafin jiki (misali, zafin fata na iya zama ƙasa da na tsokar kwarangwal. ).Ga al'adar tantanin halitta, yawan zafi yana da matsala mafi tsanani fiye da zafi.Saboda haka, yawan zafin jiki a cikin incubator yawanci ana saita shi kaɗan ƙasa da mafi kyawun zafin jiki.
2.4.1 Mafi kyawun zafin jiki don layin salula daban-daban
Yawancin layukan tantanin halitta na mutane da na dabbobi masu shayarwa ana kiyaye su a 36°C zuwa 37°C don ingantaccen girma.
Kwayoyin kwari suna noma a 27 ° C don ingantaccen girma;suna girma a hankali a ƙananan zafin jiki da yanayin zafi tsakanin 27 ° C da 30 ° C.Sama da 30 ° C, ƙarfin ƙwayoyin kwari yana raguwa, ko da ya koma 27 ° C, ƙwayoyin ba za su dawo ba.
Layukan salula na Avian suna buƙatar 38.5°C don isa iyakar girma.Ko da yake ana iya kiyaye waɗannan ƙwayoyin a 37 ° C, za su yi girma a hankali.
Layukan salula da aka samu daga dabbobi masu jin sanyi (kamar amphibians, kifin ruwan sanyi) na iya jure yanayin zafi mai faɗi daga 15°C zuwa 26°C.
Lokacin aikawa: Fabrairu-01-2023