Immediately after resusution, the cell condition is poor and some dead cells may appear. The dead cells can be removed by changing the medium overnight or the next day, or by taking 1ml from the original dish to a new culture dish while changing the medium. The amount of serum in the culture medium can be appropriately increased.

SP2/0 is a semi-adherent cell, which is round and transparent, presenting a string-like appearance. Generally, a 1ml pipette tip is sufficient for blowing and suspending.

SP2/0 cells are sourced from antibody drug enterprises and have a very good effect when used as antibodies. Most antibody drug enterprises do not have a clear demand for the preparation of hybridoma. After they complete the hybridoma treatment, they need to develop genetic engineering antibodies and then express them using cells such as CHO. They must have clear authorization and clear genetic information for this CHO and other cells. We do not provide genetic information and do not offer authorization.

Changing the culture medium may lead to unsuccessful cell culture. Upon receipt, it is necessary to use the same RPMI-1640 medium as the original product (Gibco?). For 11875) Medium, after completing the cell culture passage and cryopreservation, you can try to replace it with another medium. When obtaining the sp20 cell line, whether in cryopreservation or resuscitation state, it is necessary to first cultivate the cells to a better state in 1640 medium. Then, if necessary, a semi-medium change method can be adopted (first 50%1640+50%DEME, then 25%1640+75%DEME, and finally 100%DEME). Gradually switch to DEME medium.

8-AG (also known as 8-nitroguanine) is a basic cytotoxic factor. The synthesis of cellular DNA mainly involves two pathways: the primary pathway and the remedial pathway. The HGPRT enzyme (i.e., hypoxanthine-guanine ribosyltransferase) is the primary synthase for purines in the remedial pathway. Under normal circumstances, cells can synthesize DNA through both of these pathways simultaneously. 8-AG is also a purine and can naturally be regarded as a raw material for DNA synthesis. However, since it is toxic, its presence can kill cells. Myeloma cells with HGPRT enzyme deficiency cannot synthesize DNA through the salvage pathway, and thus can survive. Therefore, all sp2/0 cells screened by 8-AG were HGPRT enzyme-deficient strains.

刚复苏后细胞状态差且会出现部分死细胞，隔夜或隔天换液去除死细胞，或者换液的同时从原皿中取1ml到新的培养皿中培养，可适当提高培养基血清用量。

SP2/0为悬浮细胞，细胞圆形透亮，呈现葡串状。一般用1ml枪头吹悬即可。

sp2/0细胞来源自抗体药物企业，做抗体的效果非常好。 一般抗体药物企业，对杂交瘤制备这块没有明确的需求，他们做完杂交瘤后，要去做基因工程抗体，然后用CHO等细胞表达；他们对这个CHO等细胞必须要有明确的授权，明确的遗传信息。 我们不提供遗传信息且不提供授权。

换用培养基，可能导致细胞培养不成功。收到后必须用和原产品培养基一致的RPMI-1640（Gibco 11875）培养基，做好细胞培养传代冻存后，可以尝试更换其他培养基。 拿到sp20细胞株，无论是冻存状态还是复苏状态，都必须先用1640培养基把细胞培养到较好的状态，然后有需要可以采用半换液的方式（首先50%1640+50%DEME，然后25%1640+75%DEME，最后100%DEME），逐步换到DEME培养基。

There is no need for frequent rejuvenation. To maintain the genetic stability of HGPRT-SP2/0 cells, 8-nitroguanine can be added for screening every three months or so after continuous culture. The reference dosage of 8-aza-guanine is 0.00013M, or you can refer to the recommended dosage in the product manual on the market. This reagent has certain toxicity. A small amount of cell death is a normal phenomenon. If there is a large amount of death, further dilution and testing are required. Working principle: 8-Azaguanine is a purine analogue that can inhibit the proliferation of microorganisms, viruses, and tumor cells. In cells containing HGPRT enzyme (i.e., hypoxanthine-guanine phosphoribosyltransferase), 8-Azaguanine is utilized by the cells as a raw material for DNA synthesis. However, since it is toxic, it can cause the death of HGPRT+ cells, while HGPRT- cells can survive. Common HGPRT-fusion chaperone cells such as SP2/0, FO, Ag14, NS1, etc., can survive in media containing 8-aza-guanine. Therefore, 8-aza-guanine is often used for screening and optimization to ensure that these cells remain sensitive to HAT screening media. Thus, the growth of partner cells that did not participate in the fusion after cell fusion can be effectively inhibited.

The SP20 cell line is a tumor cell line that can proliferate infinitely and has the characteristic of immortalization. It is no longer restricted by the number of normal cell proliferations. Therefore, the actual generation information of the cells has become blurred, and the traditional concept of cell algebra is no longer applicable. Theoretically, they can proliferate continuously. However, in practice, to ensure cell quality, we will record and monitor the quality of the cell culture process. Moreover, we have a large customer base and our technical services are continuously using this cell line, so we can ensure that this cell can meet the requirements of cell fusion. In addition, we have a complete technical support and after-sales service system. If you encounter any quality issues after receiving it, you can contact us at any time.

Generally, 1640 medium is contained, but sometimes it may degrade during the storage process. For safety's sake, we suggest adding it