Hematopoietic Stem Cell Sorting via CD110 CD135 APLNR Markers
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Solution Overview
Problem
Current methods for obtaining hematopoietic stem cells for transplantation often result in insufficient cell numbers and reduced multilineage and engraftment potentials, particularly with non-hematopoietic sources, which limits the efficiency of allogeneic hematopoietic cell transplantation.
Innovation Solution
An in vitro method involving the sorting of hematopoietic stem cells based on the expression of CD135, CD110, and apelin receptor (APLNR) to enrich for cells capable of long-term multilineage engraftment and self-renewal, using a specific liquid culture medium that includes factors like SCF, TPO, FLT3-L, BMP4, VEGF, IL3, IL6, IL1, GCSF, and IGF1 for differentiation of pluripotent stem cells into the endo-hematopoietic lineage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If hematopoietic stem cells are obtained from non-hematopoietic sources or through conventional protocols, then the availability of cell sources is expanded, but the cell numbers remain insufficient and engraftment potential is reduced
Solution Approach 1:
The invention changes the phenotypic parameters used for cell selection from conventional markers (CD34+, CD45+) to a novel marker combination (CD110+ CD135+ APLNR+). This parameter change enables the isolation of cells with superior engraftment potential while expanding the available cell numbers from non-hematopoietic sources such as induced pluripotent stem cells and cord blood.
Solution Approach 2:
The invention copies the successful engraftment characteristics of native hematopoietic stem cells by selecting for a specific phenotypic profile (CD110+ CD135+ APLNR+) in differentiated cells from non-hematopoietic sources. This copying approach allows cells from alternative sources to acquire the functional properties needed for successful transplantation.
2Reliability
If cell sorting is performed to enrich for transplantable HSCs, then engraftment potential is improved, but the number of recoverable cells is reduced
Solution Approach 1:
The invention uses a multi-marker phenotypic profile (CD110+ CD135+ APLNR+) rather than single-marker sorting to define transplantable HSCs. This parameter change enables more precise enrichment that maintains higher cell numbers by identifying a specific functional subset rather than excluding all non-HSCs.
Solution Approach 2:
The invention applies partial sorting based on the CD110+ CD135+ APLNR+ phenotype to enrich for cells with engraftment potential while retaining sufficient cell numbers for transplantation. This partial enrichment approach balances the need for improved engraftment with the need to maintain adequate cell doses.
3Ease of operation
If conventional HSC markers are used for sorting, then the process is simple, but the isolated cells show poor engraftment potential
Solution Approach 1:
The invention changes the sorting parameters from conventional HSC markers to a novel combination (CD110+ CD135+ APLNR+). Although this increases the complexity of the sorting process, it dramatically improves engraftment potential by identifying cells with the correct functional phenotype rather than just structural characteristics.
Solution Approach 2:
The invention replaces the conventional mechanical/physical sorting approach based on traditional markers with a biologically-functional sorting strategy using the CD110+ CD135+ APLNR+ phenotype. This substitution enables the isolation of cells with proven engraftment capability rather than relying on historical marker associations.
Data Source
AI summary
The present invention relates to a method of preparing hematopoietic cell graft or enriching a population of cells for hematopoietic stem cells that are capable of long-term multilineage engraftment and self-renewal. It also relates to hematopoietic grafts comprising said hematopoietic stem cells as well as their uses in therapy.