Renal Cell Aggregates With Enhanced Transporter Gene Expression
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Solution Overview
Problem
Conventional renal cell aggregates have insufficient expression levels of transporter genes necessary for evaluating pharmacokinetics and toxicity, limiting their effectiveness in drug discovery and kidney-related disease research.
Innovation Solution
A renal cell aggregate with enhanced expression levels of OAT1, OAT3, OCT2, MATE1, MATE2, MDR1, and URAT1 genes, and a method for producing such aggregates through pre-culture, aggregate formation, and standing steps, ensuring optimal conditions for gene expression and cell function.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional renal cell aggregate is used, then it can be obtained through simple culture methods, but the expression level of transporter gene is insufficient
Solution Approach 1:
The patent applies preliminary action by pre-culturing renal cells under specific conditions (seeding density of 2.1×10³ cells/cm², passage number 3 or 4, doubling time 20-36 hours) before aggregate formation. This pre-culture step prepares the cells with optimal characteristics for subsequent aggregate formation, ensuring high transporter gene expression without complicating the overall process. The pre-cultured cells are then harvested and used to form aggregates, which are finally allowed to stand for 24 hours or more to achieve mature functional aggregates with enhanced gene expression.
2Productivity
If renal cells are cultured at high seeding density, then aggregate formation is faster, but gene expression level decreases
Solution Approach 1:
The patent applies parameter changes by optimizing the seeding density to a specific range (2.1×10³ cells/cm²) and controlling cell passage number (3 or 4) and doubling time (20-36 hours). These parameter optimizations ensure that cells are in the optimal state for both aggregate formation and maintaining high transporter gene expression. The specific parameters balance the competing requirements of formation speed and gene expression, achieving both objectives simultaneously rather than requiring a trade-off.
Data Source
Figure 1
AI summary
Provided is a renal cell aggregate in which an expression level of a transporter gene is improved. One aspect of the present invention is a renal cell aggregate. The aggregate satisfies at least one or more of the following conditions: (a) an expression level of OAT1 gene is 1 × 10-3 or more (GAPDH ratio); (b) an expression level of OAT3 gene is 1 × 10-4 or more (GAPDH ratio); (c) an expression level of OCT2 gene is 1 × 10-3 or more (GAPDH ratio); (d) an expression level of MATE1 gene is 1 × 10-4 or more (GAPDH ratio); (e) an expression level of MATE2 gene is 1 × 10-5 or more (GAPDH ratio); (f) an expression level of MDR1 gene is 1 × 10-3 or more (GAPDH ratio); and (g) an expression level of URAT1 gene is 1 × 10-6 or more (GAPDH ratio).