Placenta-Derived Exosome Isolation Using Segmented Bioreactors
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Solution Overview
Problem
Current methods for isolating and utilizing exosomes for therapeutic purposes face challenges in producing large quantities and identifying specific populations with distinct markers for effective disease treatment.
Innovation Solution
The development of methods to produce, isolate, and characterize placenta-derived exosomes with specific markers such as CD1c, CD20, CD24, CD25, CD29, and others, which can be identified using flow cytometry, for use in treating various diseases by administering them to patients.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional exosome isolation methods are used, then exosomes can be obtained, but large quantities cannot be produced efficiently
Solution Approach 1:
The placenta is divided into multiple segments or portions that can be cultured simultaneously in parallel bioreactors. This segmentation allows for scalable production where multiple placental segments contribute to exosome generation, thereby increasing overall productivity without proportionally increasing isolation time for each individual segment.
Solution Approach 2:
Placenta segments are pre-cultured in bioreactors to allow exosomes to accumulate in the culture medium before isolation is initiated. This preliminary action of pre-culturing enables larger quantities of exosomes to be produced in advance, reducing the actual isolation time when therapeutic quantities are needed.
2Manufacturing precision
If exosomes are isolated without specific marker identification, then isolation process is simpler, but specific therapeutic populations cannot be distinguished
Solution Approach 1:
Flow cytometry utilizes fluorescently labeled antibodies that bind to specific surface markers on exosomes. Different markers are detected through distinct fluorescent colors, enabling visual differentiation and precise identification of specific exosome populations based on their marker expression profiles without requiring complex manual sorting procedures.
Solution Approach 2:
Fluorescently labeled antibodies serve as intermediaries between the detection system and exosome surface markers. These antibody intermediaries specifically bind to target markers (such as CD9, CD63, CD81, or placenta-specific markers) and translate the presence/absence of specific markers into detectable fluorescent signals, enabling precise population identification.
3Reliability
If placenta-derived exosomes are used, then therapeutic benefits are achieved, but isolation from other sources cannot be distinguished
Solution Approach 1:
The exosome population is characterized by its unique local quality - specific surface marker expression profile that is distinctive to placental origin. Rather than relying on generic exosome markers, the method identifies placenta-derived exosomes through their specific marker pattern (such as co-expression of CD9, CD63, CD81 with placenta-specific markers), ensuring therapeutic reliability by confirming origin.
Solution Approach 2:
Flow cytometry provides feedback on the exosome population's marker expression profile, allowing verification of placental origin before therapeutic administration. This feedback mechanism ensures that only exosomes with the correct origin-specific marker pattern are selected for therapy, maintaining reliability while preventing use of exosomes from incorrect sources.
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
These methods enable the effective isolation and administration of placenta-derived exosomes, promoting angiogenesis, immune modulation, and tissue repair, demonstrating potential in treating a range of diseases including lung, liver, kidney, and cardiovascular disorders.
Implementation Method 1
The exosomes described herein can be identified by their morphology and/or molecular markers
Implementation Method 2
such exosomes are positive for one or more markers, e.g., as determinable by flow cytometry, for example, by fluorescence-activated cell sorting (FACS)
Data Source
AI summary
The present invention provides method of treating diseases, disorders and conditions in a human subject comprising administering to the subject a population of exosomes or a composition comprising a population of exosomes, wherein said population of exosomes is positive for CD1c, CD20, CD24, CD25, CD29, CD2, CD3, CD8, CD9, CD11c, CD14, CD19, CD31, CD40, CD41b, CD42a, CD44, CD45, CD49e, CD4, CD56, CD62P, CD63, CD69, CD81, CD86, CD105, CD133-1, CD142, CD146, CD209, CD326, HLA-ABC, HLA-DRDPDQ, MCSP, ROR1, SSEA-4, or combinations thereof. Such diseases, disorders and conditions include lung, liver, central nervous system, kidney, cardiovascular, gastrointestinal, spleen, eye, systemic and ageing associated diseases, disorders, and conditions.