Cell Culture Medium with Hemoglobin for Tumor Cell Survival
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Solution Overview
Problem
Current cell culture media struggle to maintain the survival of primary human cells, particularly tumor cells, due to a lack of critical growth and survival factors, leading to unreliable drug sensitivity assays and limited ability to test multiple drug combinations, with conventional media often causing overestimation of drug efficacy and incompatibility with certain drugs like fluorescent anthracyclines.
Innovation Solution
A cell culture medium based on anti-coagulated total blood with hemoglobin levels between 8 to 16 g/dl, where cells are disrupted and insoluble remnants are removed, preserving in vivo conditions and incorporating low molecular weight nutrients and reduced glutathione to support extended survival and high-throughput drug sensitivity testing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional cell culture media are used, then the media can be produced in large quantities at low cost, but they lack critical growth and survival factors needed to maintain primary human cells
Solution Approach 1:
The patent uses serum-free protein supplements as intermediary substances that contain the necessary growth and survival factors. These supplements act as mediators between the simple chemically-defined base medium and the complex requirements of primary human cells, providing essential factors without using traditional serum-based media.
Solution Approach 2:
The culture medium is formulated as a composite system combining chemically-defined components with serum-free protein supplements. This composite approach integrates the simplicity of defined media with the biological activity of protein-containing supplements, creating a medium that supports primary human cell growth without the complexities of traditional serum-based formulations.
2Reliability
If primary human tumor cells are cultured in conventional media, then the culturing process can be simplified, but the cells fail to survive and proliferate due to lack of survival factors
Solution Approach 1:
Serum-free protein supplements serve as intermediaries that deliver essential survival factors to primary human tumor cells. These supplements bridge the gap between the simplified culture environment and the complex biological needs of tumor cells, enabling their survival and proliferation without requiring complex serum-based media or specialized culture conditions.
3Measurement precision
If conventional cell culture media are used for drug sensitivity assays, then the assays can be performed with standard protocols, but they overestimate drug efficacy due to lack of survival factors
Solution Approach 1:
The serum-free protein supplements act as intermediaries that create a more physiologically relevant culture environment, thereby improving the accuracy of drug sensitivity measurements. By providing necessary survival factors, these supplements enable tumor cells to maintain their in vivo-like responses to drugs, eliminating the overestimation of drug efficacy that occurs in conventional media.
4Measurement precision
If fluorescent anthracyclines are used in conventional media, then drug fluorescence can be detected, but the media cause autofluorescence that interferes with accurate measurement
Solution Approach 1:
The invention extracts and eliminates the source of autofluorescence by using serum-free protein supplements instead of conventional serum-based media. This removal of autofluorescent components from the culture medium eliminates the interference with fluorescent anthracycline detection, thereby improving measurement precision.
Data Source
AI summary
The present invention relates to a cell culture medium for culturing human cells comprising an anti- coagulated total blood material wherein the hemoglobin level is from about 8 to about 16 g/dl. More particularly, the invention provides a cell culture medium in which cells present in the blood are disrupted and the insoluble remnants of the lysated cells are removed. Further, the invention provides a method for the preparation of a cell culture medium for culturing human cells, according to the invention.