1. Current techniques to culture cancer cells from patient tumors, types of tumors that can be cultivated in vitro
1.1. Some examples of tumors that can be cultivated in vitro are Gallbladder carcinoma, Mesothelioma, Osteogenic carcinoma, Ovary carcinoma, Liposarcoma, Kideny carcinoma, Colon adenocarcinoma, Nasal septum anaplastic carcinoma.
1.1.1. One approach to culturing cancer cells from patient tumors is to test the chemosensitivity of cancer cells obtained from the patients tumor. 3D structure represents a promising method for modeling patient tumors in vitro.
1.1.1.1. The aim of of this technique was to evaluate how closely short-term spheroid cultures of primary colorectal cancer cells resemble the original tumor.
1.1.1.1.1. (PDAC) or Pancreas development and primary human pancreatic adenocarinoma is another technique to culture cancer cells in vitro.
2. DNA and RNA sequencing of cancer cells from tumors-
2.1. One method of sequencing DNA and RNA is using clone align to accurately assign a cell's gene expression profile to its clone-of-origin.
2.1.1. Clone align is also applied to breast cancer xenografts and high-grade ovarian cancer cell lines; and discover biologic pathways not visible using only DNA and RNA sequencing.
2.2. The study in the attached article concluded that they were able to create a statistical framework that would allow them to assign measured cells into cancer cell clones. This type of RNA and DNA sequencing would allow advanced studies and testing towards treatments for cancer.
2.2.1. Campbell, K. R., Steif, A., Laks, E., Zahn, H., Lai, D., McPherson, A., Farahani, H., Kabeer, F., O'Flanagan, C., Biele, J., Brimhall, J., Wang, B., Walters, P., Consortium, I., Bouchard-Côté, A., Aparicio, S., & Shah, S. P. (2019). clonealign: statistical integration of independent single-cell RNA and DNA sequencing data from human cancers. Genome Biology, 20(1), NA. https://link.gale.com/apps/doc/A581445430/AONE?u=temple_main&sid=AONE&xid=d135f6a5
3. Use of animal models (mice, zebra fish, avatar models) for implanting patient tumor cells to test anti-cancer drugs
3.1. A study on ovarian cancer was done in England where they had 3 control groups of mice - 1 treated with itraconazole, 1 treated with paclitaxel, and 1 treated with both itraconazole and paclitaxel.
3.1.1. Itraconazole is a widely used antifungal drug that has shown to have effects as an anti-cancer drug.
3.1.1.1. When the mice with the patient derived tumor cells were treated with the paclitaxel and itraconazole combination, it showed a decrease in tumor weight than the mice who were treated by paclitaxel alone or itraconazole alone.
3.1.1.1.1. Choi, C. H., Ji-Yoon, R., Young-Jae, C., Jeon, H., Jung-Joo, C., Ylaya, K., . . . Jeong-Won, L. (2017). The anti-cancer effects of itraconazole in epithelial ovarian cancer. Scientific Reports (Nature Publisher Group), 7, 1-10.
3.2. A study on glioblastoma was done where they tested dexamethasone on ex vivo models - mouse retina, mouse brain slices, pig brain slices, and human brain slices. They used mouse retina to see if it would be a good substitute for brain tissue - and it was.
3.2.1. Dexamethasone is a drug that passes the blood-brain-barrier and has great bioavailability. It has shown effects in reducing edema in the brain.
3.2.1.1. First, they tested to see if human glioblastoma cells would replicate on these ex vivo models and found that they did. They treated them with dexamethasone and saw that it reduced dispersion to other parts. However, it happened slower on the human brain than it did on the pig and mouse.
3.2.1.1.1. Then, they tested to see if it was dose specific on the mouse retina and mouse brain slices. They found that if dexamethasone was removed, then the dispersion would occur. They would have to keep dexamethasone timed accordingly so that it is continuously working. Since the retina is a neural tissue and it worked on stopped dispersion at a low dose, it shows that it can work on human tumor cells as well and is a good neural substrate.
4. 3D culture of patient tumor cells for drug testing
4.1. Two-dimensional (2D) cell culture is a valuable method for cell-based research but can provide unpredictable, misleading data about in vivo responses. Three-dimensional (3D) cell environments mimic tumor characteristics and cell-cell interactions to better characterize the tumor formation response to chemotherapy.
4.1.1. A study was done constructing a 3D cell scaffold using gelatin methacryloyl (GelMA) and then comparing cell survival in 3D and 2D cell cultures. Cell-cell interactions were measured by measuring e-cadherin and n-cadherin secreted via the epithelial-mesenchymal transition (EMT). This study was done with bladder cancer cells
4.1.1.1. 3D cell cultures showed higher cancer cell proliferation rates than 2D cell cultures, and the 3D cell culture environment showed higher cell-to-cell interactions through the secretion of E-cadherin and N-cadherin.
4.1.1.1.1. These results show that 3D human cell culture systems can recapitulate the tumor heterogeneity and allow for better understanding of the molecular mechanisms that control cell-to-cell interactions.This technique can be used to create a cancer cell-like environment for a drug screening platform.
4.2. Photodynamic therapy of cancer (PDT) is a cancer treatment modality which utilizes a photoactivatable drug (photosensitizer, PS), light of the appropriate wavelength ensuring PS activation, and interstitial molecular oxygen.
4.2.1. In a previous study, scientists studied Cercosporin (Cerco), a naturally derived PS on 2D cultures but then conducted a more recent study done on 3D cultures. The scientists then compared the results of each study.
4.2.1.1. In their first study, they found Cerco to be a very potent PS, effectively killing all the cell lines at different light doses applied. The recent study on the effects of Cerco in 3D spheroid cultures was an extension of the previous study. Both studies showed Cerco to be a very potent PS but the 3D structures more closely mimic tumors and allow for better understanding and research.
4.2.1.1.1. The majority in-vitro studies have been conducted in cell mono-layers looking at cell–cell interactions where 3D structural considerations do not apply. It is therefore very important, especially for PDT applications where the light penetration into tissue is limited, to use 3D cultures which are structurally more similar to solid tumors.
5. Companies or institutions doing ex vivo culture of patient cancer cells for drug matching
5.1. Ex vivo explant assay has been used to access drug response using cyro-preserved ovarian tissue
5.1.1. The ex vivo tissue explant assay maintained viable tumor cells in an intact tumor microenvironment similar to the in vivo
5.1.1.1. The combination of 4-MU and Carboplatin significantly increased apoptosis and reduced proliferation in chemoresistant tissues
5.1.1.1.1. The ex vivo explant assay that is robust and cost effective was successfully used in assessing the sensitivity to 1st line chemotherapy drugs and for screening potential novel therapeutics
5.2. Dendritic cell (DC) vaccination has been studied extensively as active immunotherapy in cancer treatment
5.2.1. The cancers this is being used on is melanoma, leukemias, brain tumors, prostate cancer, renal cell carcinomas, pancreatic cancers and several others
5.2.1.1. Using clinical testing of ex vivo generated mRNA-transfected DCs, with choice of tumor antigens and RNA-source, and the design of better DCs for vaccination by transfection of mRNA-encoded functional proteins allows us to purposeful manipulate the DCs’ phenotype and function to enhance their immunogenicity.