Animal Origin-Free Cell Sheet Culture Supernatant
Solution Overview
Problem
Current methods for preparing mesenchymal stem cell sheets face challenges due to the use of animal-derived components and exogenous growth factors, which pose safety risks and limit the therapeutic effectiveness of the cell sheets and their culture supernatants.
Innovation Solution
A method involving a sheet-forming medium with simple ingredients, including a basal medium and human serum albumin as a binder, without animal-derived components or exogenous growth factors, and using human fibrinogen as a coating matrix to promote cell adhesion and sheet formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If serum-containing medium is used to promote cell attachment and adhesion, then cell sheet formation is improved, but safety is compromised due to bovine serum albumin and heterologous macromolecules
Solution Approach 1:
The patent extracts and removes the harmful bovine serum albumin and heterologous macromolecules from the culture medium while retaining the essential function of promoting cell attachment and adhesion. This is achieved by using a purified human-derived binder that selectively provides the adhesion-promoting properties without the allergenic components.
Solution Approach 2:
The patent introduces a human-derived binder as an intermediary substance that mediates between the cell and the culture medium. This binder serves as a safe alternative to bovine serum, providing the necessary adhesion function while eliminating the harmful allergenic components through human origin and purification.
2Productivity
If exogenous growth factors are added to support cell growth in serum-free medium, then cell proliferation is improved, but safety is compromised due to residues posing allergic risks
Solution Approach 1:
The patent removes exogenous growth factors from the culture medium while maintaining cell proliferation capability. This is achieved by using a purified human-derived binder that naturally supports cell growth without requiring additional exogenous growth factors, thereby eliminating the risk of residues causing allergic reactions.
Solution Approach 2:
The patent enables the culture medium to support cell proliferation through its own inherent components, specifically the purified human-derived binder, rather than relying on externally added growth factors. This self-sufficient approach eliminates the need for exogenous substances that could pose safety risks.
3Reliability
If scientific research grade reagents such as fibronectin and laminin are used as coating matrix, then cell adhesion is improved, but GMP compliance is compromised
Solution Approach 1:
The patent extracts and removes the non-GMP-compliant scientific research grade reagents (fibronectin and laminin) from the coating matrix while retaining the essential cell adhesion function. This is achieved by using a purified human-derived binder that provides equivalent or superior adhesion properties and meets GMP compliance requirements.
Solution Approach 2:
The patent changes the origin and purity parameters of the coating matrix from scientific research grade (non-GMP) to pharmaceutical grade (GMP-compliant). This parameter change maintains the cell adhesion function while ensuring compliance with good manufacturing practice standards for clinical applications.
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
The resulting mesenchymal stem cell sheets are safer, more durable, and capable of secreting high levels of pro-angiogenic and anti-inflammatory factors, enhancing their therapeutic potential for treating autoimmune diseases and tissue damage.
Implementation Method 1
using human fibrinogen as a coating matrix to promote cell adhesion
Implementation Method 2
human serum albumin as a binder
Data Source
AI summary
The present disclosure provides an animal origin-free cell sheet culture supernatant, for example, a human umbilical cord mesenchymal stem cell sheet culture supernatant and a preparation method for the animal origin-free cell sheet culture supernatant. The present disclosure further provides use of the animal origin-free sheet culture supernatant, for example, a human umbilical cord mesenchymal stem cell sheet culture supernatant, in the prevention and/or treatment of a skin disorder, the treatment of an autoimmune system disease, and the damage repair of a damaged tissue.