什么是無(wú)血清MSC培養(yǎng)基？
發(fā)布時(shí)間：
2022-12-24
作者：
培養(yǎng)基生產(chǎn)公司
近幾年，隨著細(xì)胞治療的進(jìn)展，對(duì)細(xì)胞體外培養(yǎng)的關(guān)注熱度也在不斷提高，作為藥品監(jiān)管的細(xì)胞制品，跟所有的藥品一樣，其評(píng)價(jià)標(biāo)準(zhǔn)也是“安全性、有效性”。因此培養(yǎng)MSC細(xì)胞的培養(yǎng)基，就成為了研究、應(yīng)用MSC細(xì)胞的企業(yè)、科研機(jī)構(gòu)的核心關(guān)注點(diǎn)。
“無(wú)血清”MSC培養(yǎng)基，是指某種培養(yǎng)基可以培養(yǎng)MSC細(xì)胞，不需額外添加任何組分，并且這種培養(yǎng)基中沒(méi)有添加血清。
市場(chǎng)上的“無(wú)血清”產(chǎn)品，可分為三類。第一種是“假的無(wú)血清”；第二種是“揣著明白裝糊涂的無(wú)血清”；第三種是“真正的無(wú)血清”。以下就簡(jiǎn)要介紹下這三種產(chǎn)品的歷史由來(lái)及如何簡(jiǎn)單的去區(qū)分。
“假的無(wú)血清”MSC培養(yǎng)基，產(chǎn)品形式一般為1瓶450ml，培養(yǎng)一瓶50ml的添加劑。銷售方稱之為無(wú)血清添加劑，但其是貨真價(jià)實(shí)的血清。做過(guò)細(xì)胞實(shí)驗(yàn)的都清楚，使用基礎(chǔ)培養(yǎng)基一般都要添加10%比例的血清。這種產(chǎn)品來(lái)源于實(shí)驗(yàn)室的發(fā)現(xiàn)，由于不同批號(hào)的血清，來(lái)自不同種群的牛，血液的成分有所不同是自然的事情，有的血清中的EGF的含量相當(dāng)?shù)?，而EGF對(duì)促進(jìn)細(xì)胞生長(zhǎng)的作用又很大。所有有經(jīng)驗(yàn)的實(shí)驗(yàn)室老師一般在用血清培養(yǎng)基培養(yǎng)MSC細(xì)胞時(shí)額外添加一些EGF,會(huì)有更好的培養(yǎng)效果。一些企業(yè)在血清中額外添加了EGF，稱之為“無(wú)血清添加劑”。這種產(chǎn)品，在市場(chǎng)上比較少，也比較容易辨識(shí)。
這種產(chǎn)品，很有迷惑性。他確實(shí)沒(méi)有添加血清，因此，從字面理解，是無(wú)血清。但他加了牛血，更具體說(shuō)是牛的血小板。我們都清楚，血液籠統(tǒng)可分為血清、血漿、血小板。在10年前，有人就發(fā)現(xiàn)，通過(guò)將血小板冷凍、再融化；再冷凍、再融化這種反復(fù)凍融的形式，可促使血小板裂解，將其中的物質(zhì)釋放出來(lái)，加入培養(yǎng)基中，可以收到與血清相類似的培養(yǎng)效果。由于血小板的含量比較少，一直以來(lái)都是作為血清生產(chǎn)的下腳料，后來(lái)伴隨血清價(jià)格的上升，處理下腳料在經(jīng)濟(jì)上顯得劃算了起來(lái)。所以，就出現(xiàn)了血小板裂解物這樣的產(chǎn)品，有的公司命名為“血清替代物”。從產(chǎn)品形式來(lái)看，一般裝量為20ml，可以添加到500ml的基礎(chǔ)培養(yǎng)基中（比如DMEM/F12，a-MEM）或添加到1000ml的減血清培養(yǎng)基中。這種產(chǎn)品，在市場(chǎng)上為數(shù)不少，比較有迷惑性。確實(shí)沒(méi)有加血清，但是加了血小板。
事實(shí)上，對(duì)MSC細(xì)胞用戶不是糾結(jié)無(wú)血清本身，而是糾結(jié)安全性。安全性來(lái)自于確定性。血小板裂解物或稱為血清替代物，與血清一樣，有許多種成分都是未知的，因此從安全性來(lái)講，難以自證安全。從事細(xì)胞培養(yǎng)的人都了解，血清中已知組分有150多種，未知組分到底是什么，有多少？沒(méi)有人可以說(shuō)的清楚。也正因?yàn)檫@個(gè)原因，在蛋白藥物生產(chǎn)方面，所有的制藥企業(yè)都選用成分明確的無(wú)血清培養(yǎng)基，而沒(méi)有哪怕一家企業(yè)選用血清的，因?yàn)闊o(wú)法證明它的安全性，當(dāng)然更無(wú)法證明從中生產(chǎn)出的抗體藥的安全性了。
實(shí)際上，如果在科研領(lǐng)域單純的培養(yǎng)細(xì)胞，血清以及血清替代物都是不錯(cuò)的選擇。當(dāng)用于臨床藥物領(lǐng)域，血清及血清替代物成分不限定的先天缺陷，注定了其無(wú)法應(yīng)用在此領(lǐng)域。
“真正的無(wú)血清”是指成分有限定的。簡(jiǎn)單說(shuō)就是包含的每一種組分都有明確的化學(xué)結(jié)構(gòu)、都是已知的?；瘜W(xué)結(jié)構(gòu)明確與成分已知是此類產(chǎn)品的最大特征。也是藥品監(jiān)管部門毫無(wú)疑義可以認(rèn)可的接觸治療性細(xì)胞產(chǎn)品的培養(yǎng)基。”真正的無(wú)血清“與血清最大的區(qū)別在于，真正的是歷經(jīng)了大量的實(shí)驗(yàn)、大量的時(shí)間、大量的文獻(xiàn)研究，研發(fā)出來(lái)的。是基于對(duì)MSC細(xì)胞的理解，專門針對(duì)MSC細(xì)胞設(shè)計(jì)出來(lái)的，在保持其細(xì)胞干性同時(shí)，保持細(xì)胞較快的生長(zhǎng)速度，以及可以接受的傳代代數(shù)。用一句話來(lái)概況他們的差別就是，無(wú)血清培養(yǎng)基是研發(fā)出來(lái)的，而血清是加工出來(lái)的。在產(chǎn)品中凝聚的技術(shù)含量差別極大。
青島日水生物 / 生物谷
In recent years, with the progress of cell therapy, the heat of attention in cell culture is increasing. As the cell product of drug regulation, the evaluation standard is "safe and effective" as all drugs are. Therefore, the culture of MSC cells has become the core concern of enterprises and research institutes that study and apply MSC cells.
Many "serum-free" media have been produced in the market, with prices ranging from 1000-4000. The user is confused.
The "serum-free" MSC medium means that a medium can cultivate MSC cells without adding any additional components, and there is no serum in this medium.
In the market, serum-free MSC medium products can be divided into three categories. The first is the "false serum-free" MSC medium; the second is "a clear and serum-free serum-free" MSC medium; the third is a "real serum-free" MSC medium. The following is a brief introduction to the historical origin of these three products and how to distinguish them simply.
"Fake serum-free" MSC medium, the product form is generally 1 bottles of 450ml medium, cultivate a bottle of 50ml additives. The seller called it a serum-free additive, but it is a genuine serum. It is clear that the use of basal medium usually adds 10% percentage of serum. This product comes from laboratory findings. Because of different batches of serum, the blood components from different populations are natural, some of the serum levels of EGF are very low, and EGF plays a great role in promoting cell growth. All experienced lab teachers usually add some EGF to MSC cells when cultured in serum medium. Some enterprises have added EGF in serum, which is called "serum free additive". This product is relatively small in the market and easy to identify.
"I think I understand the serum-free" MSC medium. This product is very confusing. He did not add any serum, so it was literally understood to be serum-free. But he added cattle blood, more specifically, bovine platelets. We all know that blood can be divided into serum, plasma and platelets. 10 years ago, it was found that platelets were frozen, thawing, and then frozen and thawing in the form of repeated freeze-thaw could cause platelet lysis to release the substance, and the culture could be added to the culture medium, which could be similar to the sera. As the content of platelets is relatively small, it has always been a waste material for the production of serum. Later, with the rise of the price of serum, the disposal of the waste material is economical. So there are products like platelet lysates, and some companies are called "serum substitutes". In the form of product, the general loading is 20ml, which can be added to the base medium of 500ml (such as DMEM/F12, a-MEM) or added to the serum medium of 1000ml. This product is quite numerous in the market and is rather confusing. No serums were added, but the blood platelets were added.
In fact, users of MSC cells are not entangled without serum themselves, but are entangled with security. Security comes from certainty. Platelet lysates, or serum substitutes, have many unknown components as well as serum. Therefore, it is difficult to prove safety in terms of safety. People engaged in cell culture know that there are more than 150 known components in serum. What are the unknown components? No one can say it clearly. For this reason, in the production of protein drugs, all pharmaceutical companies choose a serum-free medium with a clear composition, and none of the companies choose a serum medium, because it is impossible to prove the safety of the medium. Of course, it is more impossible to prove the safety of the antibody produced from it.
In fact, in the field of scientific research, pure culture cells, serum and serum substitutes are good choices. When used in the field of clinical medicine, the innate defects of the serum and serum substitutes are not bound to be destined to be applied in this field.
"Real serum-free" MSC medium. It refers to a medium defined by a component. In short, each component contained in the medium has a clear chemical structure and is known. Clear chemical structure and known components are the biggest characteristics of such products. It is also a medias that the drug regulatory authorities can identify with contact with therapeutic cell products. The biggest difference between the real serum-free "and the serum-free medium" is that the real serum-free medium has been developed by a lot of experiments, a lot of time, a lot of literature research. It is based on the understanding of MSC cells, designed specifically for MSC cells, maintaining the cell's rapid growth at the same time, as well as the acceptable subgeneration algebra. In a word, the difference between them is that the serum-free medium is developed and the serum medium is processed. There is a great difference in the technical content condensed in the product.