Sertoli Cell Expansion via EGF and bFGF Culture
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Solution Overview
Problem
The lack of a reliable method for isolating and proliferating primary human Sertoli cells has limited their availability for research and therapeutic applications, such as immune protection for transplanted cells and treatment of male infertility, due to their immunosuppressive properties and role in reproductive health.
Innovation Solution
A methodology for isolating and expanding primary human Sertoli cells from adult testicular tissue, allowing them to proliferate in vitro, be frozen, and continue proliferating post-thawing, enabling the production of a large number of cells suitable for various therapeutic and research uses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used to isolate human Sertoli cells, then cell isolation is achieved, but the cells do not proliferate in culture
Solution Approach 1:
The patent changes the chemical parameters of the culture medium by adding epidermal growth factor (EGF) at concentrations of 1-100 ng/mL and basic fibroblast growth factor (bFGF) at concentrations of 1-100 ng/mL to the culture medium. This parameter change transforms the culture conditions from those that merely maintain Sertoli cell viability to conditions that actively stimulate cell proliferation, resolving the contradiction between achieving cell isolation and enabling cell proliferation.
2Quantity of substance
If a small number of Sertoli cells are isolated from testis tissue, then cell purity is maintained, but the quantity of cells is insufficient for therapeutic applications
Solution Approach 1:
The patent performs preliminary action by adding growth factors EGF and bFGF to the culture medium before the Sertoli cells are plated. This pre-preparation of the culture environment ensures that when the cells are isolated from testis tissue, they immediately encounter proliferative signals, enabling them to expand from a small initial number to sufficient quantities for therapeutic applications without requiring complex sequential interventions.
Solution Approach 2:
The patent uses growth factors EGF and bFGF that naturally stimulate Sertoli cell proliferation in vivo, effectively copying the physiological conditions that enable cell expansion. This approach allows a small number of isolated cells to be amplified into large quantities suitable for therapy, resolving the contradiction between maintaining cell purity during isolation and achieving sufficient cell quantity for applications.
3Duration of action of stationary object
If Sertoli cells are frozen for storage, then cell preservation is achieved, but cell functionality may be compromised
Solution Approach 1:
The patent performs preliminary action by establishing robust culture conditions with EGF and bFGF before freezing the cells. This ensures that cells are in a healthy, proliferative state with optimal metabolic activity before cryopreservation, which improves their ability to withstand freezing and maintain functionality after thawing. The growth factors prepare the cells for long-term storage by enhancing their overall vitality and stress resistance.
Data Source
AI summary
Technology for the isolation and propagation of primary human Sertoli cells from normal testes tissue, including cultures of proliferative primary human Sertoli cells for research and clinical applications, and a pharmaceutical composition for cell therapy, ex vivo gene therapy, and for the reduction of autoimmune, allograft, and xenograft immune reactions.