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原代細(xì)胞的取材要求

時(shí)間:2021/9/15閱讀:292
分享:
原代細(xì)胞取材的基本要求:
 
  (1)取材要注意新鮮和保鮮 新鮮組織易于培養(yǎng)成功,取材時(shí)應(yīng)盡量在4~6h內(nèi)能制作成細(xì)胞,盡快入箱培養(yǎng),若不能即時(shí)培養(yǎng),應(yīng)將組織浸泡于培養(yǎng)液內(nèi),于4℃存放。若組織塊較大,應(yīng)在清除表面壞死組織、脂肪和結(jié)締組織后,切碎于培養(yǎng)液內(nèi)4℃存放,但時(shí)間不能超過(guò)24h。對(duì)于已切碎的組織或血液、淋巴組織應(yīng)加入含10%二甲基亞砜的培養(yǎng)基,按細(xì)胞凍存方式于液氮中冷凍保存。
 
  (2)取材應(yīng)嚴(yán)格無(wú)菌 所取標(biāo)本材料應(yīng)在無(wú)菌條件下進(jìn)行,為了確保無(wú)菌,對(duì)所取材料若疑有污染的可能,應(yīng)將所取組織在含高濃度抗菌素(400單位/mL)甚加入適量的兩性霉素B或10%達(dá)克寧液的培養(yǎng)液內(nèi)于4℃下存放2小時(shí)以上,再用PBS洗2~3次,以確保所取材料無(wú)菌。要用無(wú)菌包裝的器皿或事先消毒好的帶少許培養(yǎng)液(內(nèi)含400單位/mL抗菌素)的小瓶等便于攜帶的物品來(lái)取材,所取材料應(yīng)避免接觸有毒有害的化學(xué)物質(zhì),如碘、汞等。
 
  (3)取材和原代細(xì)胞制作時(shí),要用鋒利的器械,如手術(shù)刀或剃須刀片切碎組織,盡可能減少對(duì)細(xì)胞的機(jī)械損傷。
 
  (4)要仔細(xì)去除所取材料上的血液、脂肪、壞死組織及結(jié)締組織,切碎組織時(shí)應(yīng)避免組織干燥,可在含少量培養(yǎng)液的器皿中進(jìn)行。
 
  (5)取材應(yīng)注意組織類型、分化程度、年齡等,一般來(lái)講,胚胎組織較成熟個(gè)體組織容易培養(yǎng),分化低的較分化高的組織容易生長(zhǎng),腫瘤組織較正常組織容易培養(yǎng)。取材時(shí)應(yīng)盡量選用易培養(yǎng)的組織進(jìn)行培養(yǎng)。
 
  (6)原代細(xì)胞取材時(shí)要同時(shí)留好組織學(xué)標(biāo)本和電鏡標(biāo)本。對(duì)組織的來(lái)源、部位、包括供體的一般情況要做詳細(xì)的記錄,以備以后查詢。


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