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Updated: Jul 3, 2026
Preparation of Multifunctional Silk-Based Microcapsules Loaded with DNA Plasmids Encoding RNA Aptamers and Riboswitches
Published on: October 8, 2021
Permeability changes of the cell-contained microcapsules visualized by confocal laser scanning microscope
Guojun Lv1, Zhijie Sun, Shuangyue Li
1Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
Abstract:
Microencapsulation of recombinant cells is a novel means for gene therapy. However, one of the major concerns is the relationship between the permeability of microcapsule and cell growth. Many studies have focused on the permeability of empty microcapsule, but little is known about the effect of the cell growth on the permeability of a cell-contained microcapsule. A combination of fluorescence labeled protein and confocal laser scanning microscope (CLSM) provides the information about the permeability changes during the cell growth. A decrease of membrane permeability was detected on the 14th day. Meanwhile, membrane surface protein fouling was also investigated. A significant increase of membrane surface protein content was detected on the 21st day. In order to study the effect of the permeability changes on the cell viability, the membrane of cell-contained microcapsules with different permeability was set up by incubating gel beads in poly-L-lysine for 5 and 30 min, respectively, to mimic the bovine serum albumin cutoff, and a retard of cell growth was found in 7 days' culture. These results showed that the protein fouling of the microcapsule membrane caused by the cell growth may be an important factor to influence cell viability.
Insights
Cell growth in microcapsules for gene therapy can decrease membrane permeability due to protein fouling. This fouling impacts cell viability, highlighting a critical factor for successful therapeutic applications.
Area of Science:
- Biotechnology
- Gene Therapy
- Biomaterials
Background:
- Microencapsulation of recombinant cells is a promising gene therapy approach.
- The relationship between microcapsule permeability and cell growth is poorly understood.
- Previous studies primarily focused on empty microcapsules, neglecting cell-induced changes.
Purpose of the Study:
- To investigate the impact of cell growth on microcapsule membrane permeability.
- To analyze protein fouling on the microcapsule surface during cell culture.
- To determine the effect of altered permeability on cell viability.
Main Methods:
- Utilized fluorescence-labeled proteins and confocal laser scanning microscopy (CLSM) to monitor permeability changes.
- Quantified membrane surface protein content over time.
- Created microcapsules with varying permeability by adjusting poly-L-lysine incubation times to mimic specific protein cutoffs.
Main Results:
- A significant decrease in membrane permeability was observed by day 14.
- Increased membrane surface protein fouling was detected by day 21.
- Microcapsules with altered permeability showed retarded cell growth after 7 days of culture.
Conclusions:
- Cell growth within microcapsules leads to protein fouling of the membrane.
- This protein fouling is a key factor influencing cell viability.
- Understanding and mitigating fouling are crucial for optimizing microencapsulation-based gene therapy.
