Primary Culture Method Using Stromal Framework
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Solution Overview
Problem
Current methods for primary culture of cancer cells from tumor tissues face challenges in achieving high success rates without relying on excessive growth factors or inhibitors, often resulting in cultures that do not accurately reflect in vivo conditions.
Innovation Solution
A method involving the culture of cells on a cell structure mimicking the stromal tissue, composed of layers of stromal cells such as fibroblasts, endothelial cells, and immune cells, which includes a vascular network structure, using a general culture medium without the need for excessive growth factors or inhibitors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If serum-free medium with growth factors and inhibitors is used for primary culture, then cell survival rate is improved, but culture cost increases and in vivo-like environment is compromised
Solution Approach 1:
The invention extracts and removes the stromal component from the tumor tissue, separating it from the cancer cells. The stroma is cultured separately to create a stromal framework, and then cancer cells are seeded onto this framework. This extraction allows the use of simple, inexpensive culture media while maintaining high cell survival rates, as the stromal framework provides the necessary support and microenvironment without requiring complex growth factor supplements.
Solution Approach 2:
The invention performs preliminary culturing of stromal cells to create a three-dimensional stromal framework before seeding the cancer cells. This preliminary action establishes a supportive microenvironment that mimics the in vivo stroma, enabling cancer cells to survive and grow using only simple culture media without the need for expensive growth factors or inhibitors.
2Reliability
If serum-free medium with growth factors is used for primary culture, then cell survival rate is improved, but in vivo-like signaling environment is compromised
Solution Approach 1:
The stromal framework acts as an intermediary between the cancer cells and the culture medium. It provides a natural, in vivo-like microenvironment that mediates cell-cell and cell-matrix interactions, allowing cancer cells to maintain their physiological signaling pathways without the need for artificial growth factors or inhibitors in the culture medium.
Solution Approach 2:
The invention creates a copied version of the in vivo stromal microenvironment by culturing stromal cells from the patient's own tumor tissue and forming a three-dimensional framework. This copied stroma provides authentic in vivo-like signaling cues to the cancer cells, enabling accurate reflection of in vivo behavior without requiring complex supplements in the culture medium.
3Ease of manufacture
If conventional primary culture method is used, then culture establishment is simplified, but success rate is low
Solution Approach 1:
The invention segments the tumor tissue into two distinct components: stromal cells and cancer cells. The stromal cells are isolated and cultured separately to form a three-dimensional framework, which then serves as a substrate for seeding the cancer cells. This segmentation approach simplifies the overall culture establishment process while significantly improving the success rate, as each component can be optimized independently.
Data Source
AI summary
A primary culture method in which cells contained in a tissue collected from a living body are primary cultured in vitro, in which the cells in the tissue collected from the living body are seeded and cultured on a top surface of a cell structure containing cells constituting a stroma and composed of a single layer or two or more cell layers laminated in the thickness direction.