Oocyte Differentiation by Gene Programming in 5-10 Day Culture
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Solution Overview
Problem
Existing methods require prolonged periods, often exceeding several weeks to months, to differentiate pluripotent stem cells into immature oocytes, which is inefficient for applications in infertility treatments and other large mammals.
Innovation Solution
Introduce specific genes such as FIGLA, NOBOX, LHX8, TBPL2, and optionally STAT3, SOHLH1, SUB1, and DYNLL1 into pluripotent stem cells or primordial germ cells, and culture them for a shorter period of about 5 to 10 days to induce immature oocytes, followed by co-culture with ovarian somatic cells to produce mature oocytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional in vitro culture methods are used to differentiate pluripotent stem cells into immature oocytes, then the differentiation process can be achieved, but the culture period required is prolonged (about 3 weeks or more in mice, potentially one or more years in primates and large mammals)
Solution Approach 1:
The patent applies preliminary action by introducing oocyte-specific genes (FIGLA, NOBOX, LHX8, TBPL2) into pluripotent stem cells before the differentiation process begins. This pre-programming of genetic factors accelerates the differentiation pathway, enabling immature oocytes to be obtained within 5-10 days instead of the conventional 3 weeks or more, thereby significantly reducing the culture period while maintaining differentiation efficiency
Solution Approach 2:
The patent changes the genetic parameters of pluripotent stem cells by introducing specific oocyte-related genes (FIGLA, NOBOX, LHX8, TBPL2) and culturing them under conditions that induce expression of these genes. This parameter change transforms the differentiation timeline from months to days, resolving the contradiction between maintaining differentiation capability and reducing culture time
2Reliability
If the culture period is extended to ensure complete differentiation into immature oocytes, then differentiation completeness is improved, but the time required and resource consumption increase
Solution Approach 1:
By introducing oocyte-specific genes (FIGLA, NOBOX, LHX8, TBPL2) into pluripotent stem cells before differentiation, the patent ensures that the differentiation process is pre-programmed to proceed correctly and completely. This preliminary genetic programming maintains differentiation completeness while dramatically reducing the culture time required from months to 5-10 days
Solution Approach 2:
The patent employs feedback mechanisms by monitoring the expression of oocyte-specific genes and differentiation markers during the culture process. This allows for optimization of culture conditions to ensure complete differentiation into immature oocytes within the shortened time frame, maintaining reliability while reducing duration
Data Source
AI summary
A method for inducing immature oocytes includes introducing four genes consisting of FIGLA, NOBOX, LHX8 and TBPL2, or transcripts or expressed proteins thereof, into at least one type of cell selected from the group consisting of pluripotent stem cells, epiblast-like cells and primordial germ cells. A method for producing mature oocytes includes introducing four genes consisting of FIGLA, NOBOX, LHX8 and TBPL2, or transcripts or expressed proteins thereof, into at least one type of cell selected from the group consisting of pluripotent stem cells, epiblast-like cells and primordial germ cells, and co-culturing the cell obtained after the introduction and ovarian somatic cells.