Post-natal Hemogenic Endothelial Cell Isolation via Marker Segmentation
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Solution Overview
Problem
Current methods fail to identify and isolate post-natal hemogenic endothelial cells (HECs) capable of generating hematopoietic cells, which are crucial for regenerating the hematopoietic system, especially in mammals after birth, limiting the possibility of treating hematopoietic disorders.
Innovation Solution
A method for isolating CD144 +< CD45 +< cells from post-natal tissues like the liver, spleen, bone marrow, skin, kidney, or lung, using specific markers such as CD180, CXCR2, CXCR3, CX3CR1, CCR9, GPR141, GPR174, SLAMF7, SLAMF9, ITGAL, ITGAX, and MS4A6D, to form a substantially pure population of HECs that can be used to treat immunodeficiency disorders.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to identify hematopoietic cells, then only established hematopoietic stem cells and progenitors can be detected, but the ability to identify and isolate post-natal hemogenic endothelial cells with hematopoietic potential is lost
Solution Approach 1:
The patent changes the detection parameters by introducing new surface markers (CD144, CD45, and additional markers like CD180, CXCR2, CXCR3, CX3CR1, CCR9, GPR141, GPR174, SLAMF7, SLAMF9, ITGAL, ITGAX, MS4A6D) that allow identification of post-natal HECs. This parameter change enables detection of cells that were previously undetectable with conventional methods, resolving the contradiction between measurement precision and adaptability.
2Reliability
If de-novo hematopoiesis is assumed not to occur post-natally, then the hematopoietic system cannot be regenerated after birth, but new therapeutic possibilities for treating hematopoietic disorders are limited
Solution Approach 1:
The patent identifies and isolates HECs with hematopoietic potential before they are needed for therapeutic regeneration. By establishing the presence of these cells post-natally and developing isolation methods in advance, the pathway is prepared for future therapeutic applications, resolving the contradiction between reliability of regeneration and ease of manufacturing therapy.
Solution Approach 2:
The patent introduces HECs as an intermediary cell type that bridges endothelial cells and hematopoietic stem cells. These HECs serve as a mediator that can generate hematopoietic cells post-natally, enabling regeneration without requiring de-novo hematopoiesis and making therapeutic intervention feasible.
3Manufacturing precision
If only CD144 + CD45 + cells are isolated, then a substantially pure population of HECs can be obtained, but the complexity of the isolation procedure increases due to multiple marker requirements
Solution Approach 1:
The patent segments the isolation process into distinct stages: initial isolation using core markers (CD144, CD45) to obtain HECs, and optional further enrichment using additional markers (CD180, CXCR2, CXCR3, CX3CR1, CCR9, GPR141, GPR174, SLAMF7, SLAMF9, ITGAL, ITGAX, MS4A6D). This segmentation allows users to choose the appropriate level of complexity based on their needs, resolving the contradiction between manufacturing precision and device complexity.
Data Source
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AI summary
Provided herein are methods of isolation and identification of post-natal hemogenic endothelial cells. Further provided are substantially purified populations of post-natal hemogenic endothelial cells, compositions of post-natal hemogenic endothelial cells, and methods to utilize hemogenic endothelial cells to regenerate the hematopoietic system in a subject.