Technology - Challenges 2D vs 3D

In the past, classical 2D cell cultures or immunodeficient animal models have been used to cultivate and test drugs on human cancer cell lines. Nowadays, there is increasing interest in three-dimensional (3D) cell cultures, a method with significant differences from flat cultured cells, both considering gene expressions and cell-cell interactions. Evidence suggests that high tumorigenic properties might be dependent on the occurrence of a small cell population, pointed out to be responsible for metastasis and recurrence. This population is called cancer stem cells (CSCs), hinted to have a lot of similarities with normal stem cells. CSCs are the main reason for chemotherapy failure as well as multi-drug resistance (MDR). The big advantage of a 3D culture is the possibility to investigate the CSCs population surrounded by its environment.

An example of our in perlo™ technology application

In recent years, interest in Cannabis sativa L. has grown as legislation is moving in the right direction.
Our recent publication (2022) had 3 objectives:
(I) compare three different cannabinoid extraction methods;
(Ii) identify and quantify compounds with anticancer activity on human pancreatic cells;
And (iii) study the mechanism of action for the induction of apoptosis in a pancreatic cancer cell line described for its drug resistance.

Such a mechanistic study has never been carried out with chemo-resistant human pancreatic cancer tumoroids generated in liquid beads: the use of an original 3D culture model was a crucial first step in this study so to mimic the patient physiological condition.
It made it possible to validate the effectiveness of the extract in an in vivo test as close as possible to an animal model.
Such a test had to be carried out before any other experiment, to avoid false positives often obtained in 2D biological tests.
While gemcitabine, a reference drug in the treatment of pancreatic cancer, only triggers a stop in G0/G1 cell cycle, cannabinoid extract also activates the cell signaling cascade leading to programmed cell death.

The reported results highlight the potential of natural products derived from medicinal hemp as part of co-treatment of pancreatic cancer, as they lead to an accumulation of intracellular superoxide ions, affect the potential of the mitochondrial membrane, induce the cessation of the G1 cell cycle and ultimately lead the pancreatic cancer cell to lethal apoptosis.

Classic 2D culture methods as contemporary 3D cultures in gel or drop, had many shortcomings, including a lack of reliability. I immediately understood the interest of our 3D Pearls cell screening for the pharmaceutical industry.”

Christian D. Muller, CSO @ Co-Founder