Organoid Model Building for Glioblastoma

Glioblastoma (GBM) is a common malignant tumor of the central nervous system, belonging to WHO grade IV, with rapid progression and poor prognosis, and difficult to be completely surgically resected due to its highly infiltrative nature of growth. Therapeutic strategies targeting tumor gene targets and tumor micro-environment have become a new research direction. Currently, commonly used GBM models are mainly tumor cell culture and animal models, but cell lines are difficult to mimic tumor heterogeneity, tumor microenvironment, and oxygen concentration gradients, and their limited variety is difficult to use to study the differences in individual genetic characteristics.

We offer GBM organoid model-building services

PDX models can reproduce the original tumor genomic features but may lose some copy number mutations after several passages. To address the limitations of in vitro models in the simulation of GBM growth microenvironment and therapeutic studies, Alfa Cytology can provide GBM organoid model building. We first obtained GBM monocytes by pretreatment of GBM cell lines and transplanted GBM monocytes into cortical organoids cultured for 30 days to build GBM organoid models.

Cortical organoid model

This in vitro grown cortical organoid model has a variety of neuronal cells and brain regions, and the in vivo growth and invasion of GBM is closely related to various neuronal cells. The cortical organoid model combines the advantages of three-dimensional matrix gel culture and oscillatory culture to simulate the entire process in vivo starting from embryonic stem cells, growing into the inner, middle and outer germ layers, and then differentiating into neuroepithelial tissue, neural tube structures, and cerebral cortex. The cortical organoid model can maintain the individual genotype and the expression level of functional proteins of the organ, which can be applied to human brain disease research and drug screening.

Culture of cortical organoids

• Induced pluripotent stem cells are cultured in low-adhesion suspension by whole stem cell medium.
• When they form mimics, they are switched to suspension culture in a neural induction medium.
• They were induced to differentiate into neural ectoderm and then switched to culture with neural differentiation medium, while the tissue was transferred into Matrigel matrix gel and cultured to form neuroepithelial tissue, and continued to be cultured on a shaker in an incubator until a cortical-like organ was obtained.

We constructed cortical organoids with fluorescent markers (RFP or GFP, obtained by lentiviral transfection or fluorescent dye labeling of cortical organoids), which can make the cortical organoids fluorescent and distinguish more significantly between normal and tumor cells.

GBM organoid model

GBM stem cells are fluorescently labeled and co-cultured with cortical organoids to construct a GBM organoid model. The tumor cells are made to adhere, fuse, and infiltrate inward with the cortical organoid tissue to mimic the characteristics of tumor growth.

The construction method we provide is to use cortical organoids as a growth vehicle for GBM cells to better simulate the growth and invasion process of GBM cells. By directly transplanting tumor cells into cortical-like organs, tumor cells can gradually proliferate from within and continuously invade and develop into normal cortical-like organ tissues, which are closer to the growth characteristics of clinical tumor cells. This can better mimic the clinical growth characteristics of GBM and the tumor microenvironment, and enable the in vitro culture of GBM cells, which can be better used for tumor treatment research.

The GBM organoid model constructed by Alfa Cytology is closer to the growth characteristics of clinical tumor cells, enabling the simulation of the tumor cell growth process in normal brain tissue. This allows you to dynamically observe the growth characteristics of GBM in the organoid, which can be better used in tumor treatment and drug screening studies for GBM. It can also be used to establish GBM culture systems in cortical organoids of patient origin for individualized GBM cell culture in vitro. Please feel free to contact our staff to submit a request for GBM organoid model construction services.