Hematological tumors mainly include various types of leukemia, multiple myeloma, and malignant lymphoma. Acute leukemia accounts for the eighth place in common malignant tumors, lymphoma is also in the top ten, and the incidence rate is increasing year by year. The overall incidence of multiple myeloma accounts for 10% in hematological malignancies.
The incidence of hematological tumors is urgent. Under the traditional radiotherapy, chemotherapy, bone marrow transplantation and other treatment methods, the treatment period is long, the cost is high, and the process is painful, but the cure rate is not very satisfactory. Childhood leukemia is the most common malignant tumor in children. In recent years, the number of clinical cases has increased year by year, and it has become the number one killer in children's cancer. Therefore, the treatment of blood tumors is a top priority. Research on blood tumors is indispensable for cell and animal models. How to choose these tools can help us to conduct better research, obtain positive data, publish papers, apply for funds, and climb the peak of life. Do you know?
1. Which cell model do I need? Is it easy to operate?
In the course of the research, in order to verify the function of genes, we often need to do genetic manipulation (overexpression, interference or knockout). However, due to the characteristics of suspension growth of blood tumor cells, the difficulty of genetic manipulation is greatly increased. Lentivirus is a very efficient gene manipulation tool that also performs well in most suspension cell lines. Here are some of the characteristics of common blood tumor cells and the application of lentivirus:
*More comprehensive pre-experiment information, please consult the local salesman
Suspension Cell Infection Tips↓↓↓
What if the cell MOI>>100? Jikai has a solution for difficult-to-infect suspension cells:
1. Primary version: The method of centrifugal infection is used to increase the contact with cells when the virus is infected, thereby improving the infection efficiency. If the cell culture plate is sealed, it is centrifuged at 1000g for 15min-1h in a flat-angle rotor centrifuge, and then returned to the incubator for normal culture;
2. Advanced version: Use GMP-grade lentivirus. Especially for some primary isolated cells, the effect of viral infection on cells can be reduced by using high-purity viruses, thereby increasing infection efficiency. Jikai has the only domestic GMP production workshop for genetic engineering, and the special purification process ensures the purity and titer of clinical grade lentivirus;
3. Ultimate Edition: If you are a CAR-T teacher, you can also use Jike's own patented CAR-T cell infection kit to ensure that T cell virus infection efficiency is 40%-80%.
Second, the cell experiment works well, how does the animal experiment do?
Animal models with good stability and reproducibility can play an important role in basic tumor research and anti-tumor drug screening. The animal models commonly used in blood tumors are: I subcutaneous xenograft model; II blood model. The subcutaneous xenograft model is easier to establish and it is easy to observe the growth state of the tumor dynamically. The blood model generally uses the number of days of survival as the end point of observation. The growth and infiltration of tumor cells in the bone marrow and organs of mice are detected by living imaging, pathological section and flow cytometry. The model is close to the pathophysiological status of leukemia patients, but the experiment is difficult.
Third , the in vitro and in vivo models have been selected, then what kind of lentiviral vector does my experiment need to use to manipulate the gene?
Promoters, fluorescent markers, and resistance tags are the three major factors we need to consider when choosing a carrier. Different components also require corresponding changes in the experiment. According to the example of a god figure, customizing your exclusive carrier is as simple as that!
Shanghai Jikai Gene Chemical Technology Co., Ltd. was established in 2002 and has a BSL-2 level lentivirus packaging laboratory. The virus production line has passed the ISO9001 quality management system verification, and the monthly average custom genetic virus product packaging has exceeded 1000 times. Lentiviral production uses six QC assays, viral purity fractionation and absolute quantitative detection of viral titers to ensure virus quality.
400 dedicated Jikai people, use professional to change your efficiency.
PS: Friends who want to know more about the Tumor Tools Guide can pay attention to the "Jikai Gene" WeChat public number, and more exciting articles waiting for you.
references
[1] T Zhou et al. "Downregulation of Mcl-1 through inhibition of translation contributes to benzyl isothiocyanate induced cell cycle arrest and apoptosis in human leukemia cells." Cell Death and Disease (2013) 4, e515.
[2] Xin Huang, et al. “Analysis of the expression of PHTF1 and related genes in acute lymphoblastic leukemia .†Cancer Cell International 2015 15:93.
[3] Wang, Yi, et al. "Downregulation of miR-3940-5p promotes T-cell activity by targeting the cytokine receptor IL-2R gamma on human cutaneous T-cell lines." Immunobiology 221.12 (2016): 1378-1381 .
[4] Jian Zhang, et al. “SUMOylation of insulin-like growth factor 1 receptor, promotes proliferation in acute myeloid leukemia.†Cancer Letters, 2015, 297–306.
[5] Zhen-Hua Chen, et al. “The lncRNA HOTAIRM1 regulates the degradation of PML-RARA oncoprotein and myeloid cell differentiation by enhancing the autophagy pathway .†Cell Death and Differentiation (2016), 1–13.
[6] Xiuxiu Liu, et al. " Elevated AEG-1 expression in macrophages promotes hypopharyngeal cancer invasion through the STAT3-MMP-9 signaling pathway." ncotarget (2016): 12886.
[7]Guo, Jiang-Rui, et al. "Hepatocyte growth factor promotes proliferation, invasion, and metastasis of myeloid leukemia cells through PI3K-AKT and MAPK/ERK signaling pathway." American journal of translational research 8.9 (2016): 3630 .
[8] Lixian Wu, et al. “Dual Inhibition of Bcr-Abl and Hsp90 by C086 Potently Inhibits the Proliferation of Imatinib-Resistant CML Cells.†Clinical Cancer Research 2015.
[9] Xiaobing Miao, et al. “Overexpression of TRIP6 promotes tumor proliferation and reverses cell adhesion-mediated drug resistance (CAM-DR) via regulating nuclear p27Kip1 expression in non-Hodgkin's lymphoma.†Tumor Biology 2016, 37, 1, 1369 –1378.
[10] Bihui Yang, et al. "SPAG6 silencing inhibits the growth of the malignant myeloid cell lines SKM-1 and K562 via activating p53 and caspase activation-dependent apoptosis." International Journal of Oncology, 2014, 649-656.
[11]Armen Mardiros, et al. "T cells expressing CD123-specific chimeric antigen receptors exhibit specific cytolytic effector functions and antitumor effects against human acute myeloid leukemia." BLOOD, 31 . 2013 .122.18.
In the previous issues, I have shared a summary of the in vitro and in vivo models and tools of several large cancer species. If you want to see it again, please post the following:
1. Review of in vitro and in vivo models of lung cancer research
2. Review of in vitro and in vivo models of colorectal cancer research
3. Overview of in vivo and in vitro models of gastric cancer research
4. Summary of in vitro and in vivo models of liver cancer
5. Summary of in vitro and in vivo models of pancreatic cancer research
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