細(xì)胞污染是細(xì)胞培養(yǎng)的大敵，預(yù)防和避免污染是細(xì)胞培養(yǎng)成功的關(guān)鍵。一旦輕敵就會前功盡棄，不僅浪費(fèi)時(shí)間，而且浪費(fèi)人力、物力，甚至造成無法彌補(bǔ)的損失。而且小編的童鞋也經(jīng)常向我抱怨：每到畢業(yè)季，就藍(lán)瘦，香菇是因?yàn)榧?xì)胞死了一批又一批，畢業(yè)遙遙無期啊！那么如何打一場漂亮的細(xì)胞保衛(wèi)戰(zhàn)呢？下面就跟隨小編一起來學(xué)起來吧~

*步：認(rèn)識細(xì)胞培養(yǎng)過程中有哪些污染？

細(xì)胞培養(yǎng)污染是指培養(yǎng)環(huán)境中侵入了對細(xì)胞生存有害的成分和造成細(xì)胞變異的異物。體外細(xì)胞培養(yǎng)污染可分為三類：物理性、化學(xué)性及生物性污染。

物理性污染：
物理性污染通過影響細(xì)胞培養(yǎng)體系中的生化成分，從而影響細(xì)胞的代謝。
物理性污染的來源：培養(yǎng)環(huán)境中的物理因素，如溫度、放射線、振動、輻射（紫外線或熒光）會對細(xì)胞產(chǎn)生影響。
物理性污染的現(xiàn)象：細(xì)胞、培養(yǎng)液或其他培養(yǎng)試劑暴露在放射線、輻射或過冷過熱的溫度中，可以引起細(xì)胞代謝發(fā)生改變，如細(xì)胞同步化、細(xì)胞生長受抑制，甚至細(xì)胞死亡。
化學(xué)性污染
化學(xué)性污染是一些對細(xì)胞有毒性的或?qū)?xì)胞產(chǎn)生刺激的化學(xué)物質(zhì)。
化學(xué)性污染的來源：
未純化的物質(zhì)、試劑、水、血清、生長輔助因子及儲存試劑的容器都可能成為化學(xué)性污染的來源。細(xì)胞培養(yǎng)的必需養(yǎng)分（如氨基酸）若濃度超過了合理的范圍，也會對細(xì)胞產(chǎn)生毒性。同樣，不同細(xì)胞系在*培養(yǎng)條件下對血清和緩沖液的要求是不一樣的，在培養(yǎng)中應(yīng)嚴(yán)格控制。
化學(xué)性污染的現(xiàn)象：化學(xué)性污染同樣會使細(xì)胞生長受抑制，甚至細(xì)胞死亡。
生物污染
生物污染包括比較容易發(fā)現(xiàn)的細(xì)菌、霉菌和酵母的污染，和較難發(fā)現(xiàn)的病毒、支原體和其他細(xì)胞的污染。
生物性污染的來源：
● 取材不慎或處理不當(dāng)
● 空氣塵埃
● 消毒不嚴(yán)
● 操作不當(dāng)
● 試劑尤其是血清污染
生物性污染的現(xiàn)象：
● 細(xì)菌：G+菌較多。
● 真菌：霉菌較多。
● 支原體：不易檢測。
● 病毒：由于病毒有種屬特異性，所以病毒污染的概率比較小。

第二步：做好預(yù)防污染措施
針對細(xì)胞污染的三個(gè)分類，做好如下預(yù)防措施：
A.物理性污染的預(yù)防
● 實(shí)驗(yàn)室的合理設(shè)計(jì)及建立規(guī)范的操作規(guī)程；
● 培養(yǎng)箱應(yīng)放在恒溫的環(huán)境中；
● 培養(yǎng)液及試劑應(yīng)放在固定位置，而且要注意避光，試劑周圍不能放置同位素；
● 從冰箱中取出的培養(yǎng)液應(yīng)在室溫條件中放置一段時(shí)間后再進(jìn)行使用，以避免過冷的溫度對細(xì)胞的影響。
B.化學(xué)性污染的預(yù)防
● 使用的所有物質(zhì)都應(yīng)是高純度的，同時(shí)正確配制和儲存培養(yǎng)液及試劑。
● 在配制液體和清洗容器時(shí)必須使用不含雜質(zhì)的超純水。
● 選用同一批次的血清。
● 在選用器皿時(shí)，根據(jù)被培養(yǎng)細(xì)胞的生長特性、培養(yǎng)方式和所用培養(yǎng)液的性質(zhì)來選用適合的器皿來達(dá)到實(shí)驗(yàn)的準(zhǔn)確性和可靠性。
C.生物性污染的預(yù)防
● 建立規(guī)范的無菌操作程序及各種規(guī)章制度，并嚴(yán)格執(zhí)行。
● 控制環(huán)境污染，同時(shí)注意實(shí)驗(yàn)人員的防護(hù)及無菌服的潔凈和無菌。
● 保證細(xì)胞培養(yǎng)基和器材無菌。
● 在細(xì)胞培養(yǎng)基中加入適量的抗生素。
預(yù)防是防止細(xì)胞培養(yǎng)過程中發(fā)生污染的辦法。只有提前做好預(yù)防工作，才能將發(fā)生
污染的可能性降到zui小程度。

Cell pollution is the enemy of cell culture. Preventing and avoiding pollution is the key to the success of cell culture. Once the enemy will come to naught, not only a waste of time, and a waste of manpower and material resources, and even cause irreparable loss. And little children's shoes often complain to me: every graduation season, the blue thin, letinous edodes is because of the cell dead batch after batch, graduation far away ah! So how do you fight a beautiful cell defense? Then follow the little editor to learn.

The first step: what are the pollution in the process of cell culture?

Cell culture pollution refers to the invasion of elements which are harmful to the survival of cells and the foreign bodies that cause cell mutation in the culture environment. In vitro cell culture pollution can be divided into three categories: physical, chemical and biological pollution.

Physical pollution:
Physical pollution affects the metabolism of cells by affecting the biochemical components of the cell culture system.
The sources of physical pollution: physical factors in the culture environment, such as temperature, radiation, vibration, radiation (ultraviolet or fluorescence), can affect cells.
The phenomenon of physical pollution: cells, culture fluids or other culture reagents exposed to radiation, radiation or supercooled superheated temperatures can cause changes in cellular metabolism, such as cell synchronization, cell growth inhibition, and even cell death.
Chemical pollution
Chemical pollution is a chemical substance that is toxic to cells or stimulates cells.
Sources of chemical pollution:
Containers of non purified substances, reagents, water, serum, growth cofactors and storage reagents can all be the source of chemical pollution. The essential nutrients (such as amino acids) in cell culture, if the concentration exceeds the reasonable range, can also be toxic to the cells. In the same way, the requirements of different cell lines to the serum and buffer solution are different under the best culture conditions, and should be strictly controlled in the culture.
Chemical pollution: chemical pollution can also inhibit cell growth and even cell death.
Biological pollution
Biological pollution includes contamination of bacteria, fungi, and yeast, which are easier to detect, and the contamination of viruses, mycoplasma and other cells that are more difficult to detect.
Sources of biological pollution:
Inadvertently or mishandled
Air dust
Disinfection is not strict
Improper operation
Reagents, especially serum pollution
The phenomenon of biological pollution:
Bacteria: more G+ bacteria.
Fungi: more fungi.
Mycoplasma: it is not easy to detect.
Virus: the virus has a species specificity, so the probability of virus contamination is relatively small.

The second step: do a good job of preventing pollution
According to the three categories of cell pollution, the following preventive measures are done:
Prevention of physical pollution in A.
The rational design of the laboratory and the establishment of a standardized operating procedure.
The incubator should be placed in a constant temperature environment.
The c*tion liquid and reagents should be placed in a fixed position, and the light should be avoided, and the isotopes can not be placed around the reagent.
The culture liquid extracted from the refrigerator should be used at room temperature for a period of time to avoid the effect of the supercooling temperature on the cells.
Prevention of B. chemical pollution
All substances used should be of high purity, and the culture fluid and reagents are properly prepared and stored.
Ultra pure water containing no impurities must be used in the preparation of liquid and cleaning containers.
Select the same batch of serum.
When selecting utensils, according to the growth characteristics of the cultured cells, the way of culture and the nature of the culture medium, we choose suitable containers to achieve the accuracy and reliability of the experiment.
Prevention of C. biological pollution
The establishment of standardized aseptic procedures and various rules and regulations, and strict implementation.
Control the environmental pollution, and pay attention to the protection of the laboratory personnel and the cleanliness and asepsis of the aseptic clothes.
Ensure the asepsis of cell culture medium and equipment.
An appropriate amount of antibiotics is added to the cell culture medium.
Prevention is the best way to prevent pollution in the process of cell culture. Only if the precautionary work is done ahead of time, can it happen
The possibility of pollution is minimized.

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