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High-Throughput Cancer Cell Sphere Formation for Characterizing the Efficacy of Photo Dynamic Therapy in 3D Cell Cultures.

Chen YC, Lou X, Zhang Z, Ingram P, Yoon E - Sci Rep (2015)

Bottom Line: Using the presented platform, we have successfully characterized the different responses in 2D and 3D cell culture to PDT.Although the CAFs can enhance the resistance to traditional chemotherapy agents, no significant difference in PDT was observed.The preliminary results suggest that the PDT can be an attractive alternative cancer therapy, which is less affected by the therapeutic resistance induced by cancer associated cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Electrical Engineering and Computer Science, University of Michigan, 1301 Beal Avenue, Ann Arbor, MI 48109-2122.

ABSTRACT
Photodynamic therapy (PDT), wherein light sensitive non-toxic agents are locally and selectively activated using light, has emerged as an appealing alternative to traditional cancer chemotherapy. Yet to date, PDT efficacy has been mostly characterized using 2D cultures. Compared to 2D cultures, 3D sphere culture generates unique spatial distributions of nutrients and oxygen for the cells that better mimics the in-vivo conditions. Using a novel polyHEMA (non-adherent polymer) fabrication process, we developed a microfluidic sphere formation platform that can (1) generate 1,024 uniform (size variation <10%) cancer spheres within a 2 cm by 2 cm core area, (2) culture spheres for more than 2 weeks, and (3) allow the retrieval of spheres. Using the presented platform, we have successfully characterized the different responses in 2D and 3D cell culture to PDT. Furthermore, we investigated the treatment resistance effect in cancer cells induced by tumor associated fibroblasts (CAF). Although the CAFs can enhance the resistance to traditional chemotherapy agents, no significant difference in PDT was observed. The preliminary results suggest that the PDT can be an attractive alternative cancer therapy, which is less affected by the therapeutic resistance induced by cancer associated cells.

No MeSH data available.


Related in: MedlinePlus

High-throughput sphere formation microfluidic chip.(a) Schematic showing cells settling into a microwell during loading. (b) Schematic showing cell aggregation and sphere formation 1 day after cell loading. (c) 3D schematic view of the microfluidic chip. (d) Enlarged 3D schematic of micro-wells. (e) The fabricated device that contains 1,024 micro-wells within a core area of 2 cm by 2 cm. The cells are loaded in the inlet (top right) and then flow through the microwell array to the outlet (bottom left).
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f1: High-throughput sphere formation microfluidic chip.(a) Schematic showing cells settling into a microwell during loading. (b) Schematic showing cell aggregation and sphere formation 1 day after cell loading. (c) 3D schematic view of the microfluidic chip. (d) Enlarged 3D schematic of micro-wells. (e) The fabricated device that contains 1,024 micro-wells within a core area of 2 cm by 2 cm. The cells are loaded in the inlet (top right) and then flow through the microwell array to the outlet (bottom left).

Mentions: The presented high-throughput sphere formation platform is composed of an array of 1,024 non-adherent microwells connected by single inlet and outlet (Fig. 1). Spheres are formed by aggregation of cells in each microwell (400 μm in depth) after cell loading (Fig. 1). When cells are loaded into the channel, a portion of the cells will settle down into a microwell and the rest will flow into the rest of downstream microwells (Fig. 1(a)). As the number of cells that settle in a well depends on the size of micro-well, we can easily generate larger and smaller spheres for different applications by changing the diameter of microwells. In our study, we used two different microwell diameters, 250 μm and 450 μm, respectively. To facilitate sphere formation by cell aggregation, micro-wells were coated with polyHEMA. PolyHEMA is a biocompatible hydrogel that has been used as a non-adherent coating material for suspension cell culture successfully for more than 30 years3839. As cells cannot adhere onto the polyHEMA-coated layer, they aggregate to form a sphere (Fig. 1(b)). Using the presented device (Fig. 1(c–e)), we can reliably generate 1,024 spheres at high throughput within 1 day.


High-Throughput Cancer Cell Sphere Formation for Characterizing the Efficacy of Photo Dynamic Therapy in 3D Cell Cultures.

Chen YC, Lou X, Zhang Z, Ingram P, Yoon E - Sci Rep (2015)

High-throughput sphere formation microfluidic chip.(a) Schematic showing cells settling into a microwell during loading. (b) Schematic showing cell aggregation and sphere formation 1 day after cell loading. (c) 3D schematic view of the microfluidic chip. (d) Enlarged 3D schematic of micro-wells. (e) The fabricated device that contains 1,024 micro-wells within a core area of 2 cm by 2 cm. The cells are loaded in the inlet (top right) and then flow through the microwell array to the outlet (bottom left).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495468&req=5

f1: High-throughput sphere formation microfluidic chip.(a) Schematic showing cells settling into a microwell during loading. (b) Schematic showing cell aggregation and sphere formation 1 day after cell loading. (c) 3D schematic view of the microfluidic chip. (d) Enlarged 3D schematic of micro-wells. (e) The fabricated device that contains 1,024 micro-wells within a core area of 2 cm by 2 cm. The cells are loaded in the inlet (top right) and then flow through the microwell array to the outlet (bottom left).
Mentions: The presented high-throughput sphere formation platform is composed of an array of 1,024 non-adherent microwells connected by single inlet and outlet (Fig. 1). Spheres are formed by aggregation of cells in each microwell (400 μm in depth) after cell loading (Fig. 1). When cells are loaded into the channel, a portion of the cells will settle down into a microwell and the rest will flow into the rest of downstream microwells (Fig. 1(a)). As the number of cells that settle in a well depends on the size of micro-well, we can easily generate larger and smaller spheres for different applications by changing the diameter of microwells. In our study, we used two different microwell diameters, 250 μm and 450 μm, respectively. To facilitate sphere formation by cell aggregation, micro-wells were coated with polyHEMA. PolyHEMA is a biocompatible hydrogel that has been used as a non-adherent coating material for suspension cell culture successfully for more than 30 years3839. As cells cannot adhere onto the polyHEMA-coated layer, they aggregate to form a sphere (Fig. 1(b)). Using the presented device (Fig. 1(c–e)), we can reliably generate 1,024 spheres at high throughput within 1 day.

Bottom Line: Using the presented platform, we have successfully characterized the different responses in 2D and 3D cell culture to PDT.Although the CAFs can enhance the resistance to traditional chemotherapy agents, no significant difference in PDT was observed.The preliminary results suggest that the PDT can be an attractive alternative cancer therapy, which is less affected by the therapeutic resistance induced by cancer associated cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Electrical Engineering and Computer Science, University of Michigan, 1301 Beal Avenue, Ann Arbor, MI 48109-2122.

ABSTRACT
Photodynamic therapy (PDT), wherein light sensitive non-toxic agents are locally and selectively activated using light, has emerged as an appealing alternative to traditional cancer chemotherapy. Yet to date, PDT efficacy has been mostly characterized using 2D cultures. Compared to 2D cultures, 3D sphere culture generates unique spatial distributions of nutrients and oxygen for the cells that better mimics the in-vivo conditions. Using a novel polyHEMA (non-adherent polymer) fabrication process, we developed a microfluidic sphere formation platform that can (1) generate 1,024 uniform (size variation <10%) cancer spheres within a 2 cm by 2 cm core area, (2) culture spheres for more than 2 weeks, and (3) allow the retrieval of spheres. Using the presented platform, we have successfully characterized the different responses in 2D and 3D cell culture to PDT. Furthermore, we investigated the treatment resistance effect in cancer cells induced by tumor associated fibroblasts (CAF). Although the CAFs can enhance the resistance to traditional chemotherapy agents, no significant difference in PDT was observed. The preliminary results suggest that the PDT can be an attractive alternative cancer therapy, which is less affected by the therapeutic resistance induced by cancer associated cells.

No MeSH data available.


Related in: MedlinePlus