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A library of mammalian effector modules for synthetic morphology.

Cachat E, Liu W, Hohenstein P, Davies JA - J Biol Eng (2014)

Bottom Line: Together with cell differentiation, these mechanisms allow populations of cells to organize themselves into defined geometries and structures, as simple embryos develop into complex organisms.Here we describe this library and demonstrate its use in the T-REx-293 human cell line to induce each of these desired morphological behaviours on command.Building on from the simple test systems described here, we want to extend engineered control of morphogenetic cell behaviour to more complex 3D structures that can inform embryologists and may, in the future, be used in surgery and regenerative medicine, making synthetic morphology a powerful tool for developmental biology and tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: University of Edinburgh, Centre for Integrative Physiology, Hugh Robson Building, George Square, Edinburgh, EH8 9XD UK.

ABSTRACT

Background: In mammalian development, the formation of most tissues is achieved by a relatively small repertoire of basic morphogenetic events (e.g. cell adhesion, locomotion, apoptosis, etc.), permutated in various sequences to form different tissues. Together with cell differentiation, these mechanisms allow populations of cells to organize themselves into defined geometries and structures, as simple embryos develop into complex organisms. The control of tissue morphogenesis by populations of engineered cells is a potentially very powerful but neglected aspect of synthetic biology.

Results: We have assembled a modular library of synthetic morphogenetic driver genes to control (separately) mammalian cell adhesion, locomotion, fusion, proliferation and elective cell death. Here we describe this library and demonstrate its use in the T-REx-293 human cell line to induce each of these desired morphological behaviours on command.

Conclusions: Building on from the simple test systems described here, we want to extend engineered control of morphogenetic cell behaviour to more complex 3D structures that can inform embryologists and may, in the future, be used in surgery and regenerative medicine, making synthetic morphology a powerful tool for developmental biology and tissue engineering.

No MeSH data available.


Related in: MedlinePlus

Effect of the adhesion effector on engineered T-REx-293 cells. (a) Levels of murine Cdh1 mRNA expression in cells of clone THAD1-34 (a representative clone of T-REx-293 cells carrying the adhesion module), uninduced or induced with 1 μg/mL tetracycline for 48 h; and in wild-type T-REx-293 cells treated with tetracycline, as a control, for 48 h. (b) Morphology of wild-type T-REx-293 cells and THAD1-34 cells growing with or without tetracycline for 48 h: wild-type cells with or without tetracycline and uninduced THAD1-34 cells spread across the culture substrate with only loose mutual association, whereas induced THAD1-34 cells adhere to one another strongly to produce dense islands separated by clear space. Scale bars: 100 μm.
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Fig4: Effect of the adhesion effector on engineered T-REx-293 cells. (a) Levels of murine Cdh1 mRNA expression in cells of clone THAD1-34 (a representative clone of T-REx-293 cells carrying the adhesion module), uninduced or induced with 1 μg/mL tetracycline for 48 h; and in wild-type T-REx-293 cells treated with tetracycline, as a control, for 48 h. (b) Morphology of wild-type T-REx-293 cells and THAD1-34 cells growing with or without tetracycline for 48 h: wild-type cells with or without tetracycline and uninduced THAD1-34 cells spread across the culture substrate with only loose mutual association, whereas induced THAD1-34 cells adhere to one another strongly to produce dense islands separated by clear space. Scale bars: 100 μm.

Mentions: After clone selection, T-REx-293 cells stably transfected with pTREx-TGFP-2A-Cdh1 expressed mouse E-cadherin after 48 h of 1 μg/mL tetracycline induction, as shown by RT-PCR analysis (Figure 4a). The primers used to detect levels of Cdh1 mRNA were specific for mouse Cdh1. When uninduced, cells show very little mouse E-cadherin mRNA transcription compared to induced cells, indicating a low level of expression leakage from the tetracycline transcriptional switch. Wild-type cells and T-REx-293 cells carrying the adhesion module but without induction showed only modest cell-cell adhesion: they grew in small, connected, ragged-edged clumps that formed a network across the culture plate (Figure 4b). Tetracycline itself had no effect on the morphology of wild-type T-REx-293 cultures. The same cells under tetracycline induction of Cdh1 formed much more compact, dense, smooth-edged, separate islands (Figure 4b). These features are as expected, for increased adhesion would increase ‘surface tension’ at the edges of the islands, minimizing their circumference.Figure 4


A library of mammalian effector modules for synthetic morphology.

Cachat E, Liu W, Hohenstein P, Davies JA - J Biol Eng (2014)

Effect of the adhesion effector on engineered T-REx-293 cells. (a) Levels of murine Cdh1 mRNA expression in cells of clone THAD1-34 (a representative clone of T-REx-293 cells carrying the adhesion module), uninduced or induced with 1 μg/mL tetracycline for 48 h; and in wild-type T-REx-293 cells treated with tetracycline, as a control, for 48 h. (b) Morphology of wild-type T-REx-293 cells and THAD1-34 cells growing with or without tetracycline for 48 h: wild-type cells with or without tetracycline and uninduced THAD1-34 cells spread across the culture substrate with only loose mutual association, whereas induced THAD1-34 cells adhere to one another strongly to produce dense islands separated by clear space. Scale bars: 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4255936&req=5

Fig4: Effect of the adhesion effector on engineered T-REx-293 cells. (a) Levels of murine Cdh1 mRNA expression in cells of clone THAD1-34 (a representative clone of T-REx-293 cells carrying the adhesion module), uninduced or induced with 1 μg/mL tetracycline for 48 h; and in wild-type T-REx-293 cells treated with tetracycline, as a control, for 48 h. (b) Morphology of wild-type T-REx-293 cells and THAD1-34 cells growing with or without tetracycline for 48 h: wild-type cells with or without tetracycline and uninduced THAD1-34 cells spread across the culture substrate with only loose mutual association, whereas induced THAD1-34 cells adhere to one another strongly to produce dense islands separated by clear space. Scale bars: 100 μm.
Mentions: After clone selection, T-REx-293 cells stably transfected with pTREx-TGFP-2A-Cdh1 expressed mouse E-cadherin after 48 h of 1 μg/mL tetracycline induction, as shown by RT-PCR analysis (Figure 4a). The primers used to detect levels of Cdh1 mRNA were specific for mouse Cdh1. When uninduced, cells show very little mouse E-cadherin mRNA transcription compared to induced cells, indicating a low level of expression leakage from the tetracycline transcriptional switch. Wild-type cells and T-REx-293 cells carrying the adhesion module but without induction showed only modest cell-cell adhesion: they grew in small, connected, ragged-edged clumps that formed a network across the culture plate (Figure 4b). Tetracycline itself had no effect on the morphology of wild-type T-REx-293 cultures. The same cells under tetracycline induction of Cdh1 formed much more compact, dense, smooth-edged, separate islands (Figure 4b). These features are as expected, for increased adhesion would increase ‘surface tension’ at the edges of the islands, minimizing their circumference.Figure 4

Bottom Line: Together with cell differentiation, these mechanisms allow populations of cells to organize themselves into defined geometries and structures, as simple embryos develop into complex organisms.Here we describe this library and demonstrate its use in the T-REx-293 human cell line to induce each of these desired morphological behaviours on command.Building on from the simple test systems described here, we want to extend engineered control of morphogenetic cell behaviour to more complex 3D structures that can inform embryologists and may, in the future, be used in surgery and regenerative medicine, making synthetic morphology a powerful tool for developmental biology and tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: University of Edinburgh, Centre for Integrative Physiology, Hugh Robson Building, George Square, Edinburgh, EH8 9XD UK.

ABSTRACT

Background: In mammalian development, the formation of most tissues is achieved by a relatively small repertoire of basic morphogenetic events (e.g. cell adhesion, locomotion, apoptosis, etc.), permutated in various sequences to form different tissues. Together with cell differentiation, these mechanisms allow populations of cells to organize themselves into defined geometries and structures, as simple embryos develop into complex organisms. The control of tissue morphogenesis by populations of engineered cells is a potentially very powerful but neglected aspect of synthetic biology.

Results: We have assembled a modular library of synthetic morphogenetic driver genes to control (separately) mammalian cell adhesion, locomotion, fusion, proliferation and elective cell death. Here we describe this library and demonstrate its use in the T-REx-293 human cell line to induce each of these desired morphological behaviours on command.

Conclusions: Building on from the simple test systems described here, we want to extend engineered control of morphogenetic cell behaviour to more complex 3D structures that can inform embryologists and may, in the future, be used in surgery and regenerative medicine, making synthetic morphology a powerful tool for developmental biology and tissue engineering.

No MeSH data available.


Related in: MedlinePlus