Murine Cell Depletion via CD9 Antibody Detection
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Solution Overview
Problem
The contamination of human xenografts with murine cells complicates molecular analysis and cell culture due to variable infiltration and overgrowth by mouse cells, making it difficult to achieve accurate downstream analysis and purification of human cells.
Innovation Solution
A process involving the use of antibodies specific to the murine CD9 epitope for detecting and depleting murine cells from xenografts, which includes fragmenting the xenograft into single cells, subjecting the sample to CD9-specific antibodies, and separating the bound cells using detection means like magnetic particles or fluorescent dyes, resulting in a substantial depletion of host cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional antibody combinations (CD45, MHC class I) are used to deplete mouse cells, then some mouse cells can be removed, but a subset of mouse cells remains undetected and contamination persists
Solution Approach 1:
The patent changes the detection parameter from conventional markers (CD45, MHC class I) to the CD9 epitope, which provides broader recognition of mouse cells across different tissues and cell types, thereby improving detection accuracy and reducing remaining contamination
Solution Approach 2:
The CD9 antibody serves as a universal marker for mouse cells, functioning across multiple tissue types and cell lineages where conventional markers like CD45 and MHC class I show limited efficacy, enabling comprehensive depletion of diverse mouse cell populations
2Reliability
If multiple antibody combinations are used to improve mouse cell depletion, then detection coverage increases, but the process complexity and time required for purification increases
Solution Approach 1:
The patent extracts the essential function of mouse cell detection and depletion into a single antibody targeting the CD9 epitope, eliminating the need for multiple antibody combinations and complex sequential purification steps while maintaining high reliability of mouse cell removal
3Quantity of substance
If mouse cells are not depleted from xenografts, then the xenograft can be maintained in culture, but mouse fibroblasts overgrow human tumor cells and contaminate the culture
Solution Approach 1:
The patent performs preliminary depletion of mouse cells using CD9 antibodies before initiating cell culture, preventing subsequent overgrowth by mouse fibroblasts and ensuring human tumor cells can proliferate without contamination, thereby maintaining both purity and productivity
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach effectively isolates human cells by recognizing and removing a vast majority of murine cells, including those from various organs, improving the purity of human cell samples for further analysis and culture, especially in cancer research, by achieving >99% depletion of contaminating mouse cells.
Implementation Method 1
subjecting the sample to antibodies specific for a murine CD9 epitope coupled to a detection means
Implementation Method 2
separating the bound cells using detection means like magnetic particles or fluorescent dyes
Implementation Method 3
separating the bound cells using detection means like magnetic particles or fluorescent dyes
Data Source
AI summary
A process for depleting host cells from a xenograft of human cells on a murine host is disclosed. The process includes fragmenting the xenograft, subjecting the sample to antibodies specific for a murine CD9 epitope coupled to a detection means, depleting the cell suspension from cells bound by the CD9-antibodies using the detection means, and collecting the cells not bound by the CD9-antibodies as target cells.