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High Content Imaging (HCI) on Miniaturized Three-Dimensional (3D) Cell Cultures.

Joshi P, Lee MY - Biosensors (Basel) (2015)

Bottom Line: High content imaging (HCI) is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates.Conventional HCI assays have been carried out on two-dimensional (2D) cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D) cell cultures.Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS).

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical & Biomedical Engineering, Cleveland State University, 1960 East 24th Street Cleveland, Ohio, OH 44115-2214, USA. p.joshi18@vikes.csuohio.edu.

ABSTRACT
High content imaging (HCI) is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates. Conventional HCI assays have been carried out on two-dimensional (2D) cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D) cell cultures. Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS). One attempt of making 3D cell culture amenable for HTS is to utilize miniaturized cell culture platforms. This review aims to highlight miniaturized 3D cell culture platforms compatible with current HCI technology.

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Miniaturized 3D cell culture systems for HCI. (A) Microwell platform (Adapted from Ref [90] with permission of The Royal Society of Chemistry). Overview of hydrogel microwell arrays fabrication process: (Step 1) A polydimethylsiloxane (PDMS) stamp containing an array of micropillars is cast on a silicon master. (Steps 2 and 3) Poly(ethylene glycol) (PEG) gel is cross-linked to contain complementary microwell array topography using the PDMS stamp as a template. (Step 4) Individual cells are trapped on the hydrogel surface after swelling and washing of the surface. (B) Cellular microarrayson a functionalized glass slide. A mixture of cells and hydrogel precursor is printed on a glass slide coated with poly(styrene-co-maleic anhydride) (PS-MA). Various polymer coating is done on top of the PS-MA coating to attach different hydrogels to the glass slide. Cells are encapsulated in a hydrogel matrix, forming 3D structures after gelation (which occurs via various mechanisms). (C) Cellular microarrays on a micropillar/microwell chip platform (Adapted by permission from Macmillan Publishers Ltd: Nature Communications, Ref [84]). Cells mixed with hydrogel are printed on top of the micropillar chip. After gelation, the micropillar chip containing cells encapsulated in hydrogel is sandwiched with a complementary microwell chip containing growth media or other reagents. (D) Microfluidicdevice. (i) Top view of a bilayer microfluidic chip fabricated with PDMS on top of a glass slide. Several inlet and outlet channels provide parallel access to cell suspension, growth medium and other reagents. (ii) Overview of the cell culture process in the microfluidic device: (Step 1) Bi-layer chip is fabricated with PDMS containing several channels on top of a glass slide. (Step 2) A mixture of cells and hydrogel precursor is fed from the cell inlet channel. (Step 3) A growth medium is supplied from the medium inlet channel for cell culture.
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biosensors-05-00768-f002: Miniaturized 3D cell culture systems for HCI. (A) Microwell platform (Adapted from Ref [90] with permission of The Royal Society of Chemistry). Overview of hydrogel microwell arrays fabrication process: (Step 1) A polydimethylsiloxane (PDMS) stamp containing an array of micropillars is cast on a silicon master. (Steps 2 and 3) Poly(ethylene glycol) (PEG) gel is cross-linked to contain complementary microwell array topography using the PDMS stamp as a template. (Step 4) Individual cells are trapped on the hydrogel surface after swelling and washing of the surface. (B) Cellular microarrayson a functionalized glass slide. A mixture of cells and hydrogel precursor is printed on a glass slide coated with poly(styrene-co-maleic anhydride) (PS-MA). Various polymer coating is done on top of the PS-MA coating to attach different hydrogels to the glass slide. Cells are encapsulated in a hydrogel matrix, forming 3D structures after gelation (which occurs via various mechanisms). (C) Cellular microarrays on a micropillar/microwell chip platform (Adapted by permission from Macmillan Publishers Ltd: Nature Communications, Ref [84]). Cells mixed with hydrogel are printed on top of the micropillar chip. After gelation, the micropillar chip containing cells encapsulated in hydrogel is sandwiched with a complementary microwell chip containing growth media or other reagents. (D) Microfluidicdevice. (i) Top view of a bilayer microfluidic chip fabricated with PDMS on top of a glass slide. Several inlet and outlet channels provide parallel access to cell suspension, growth medium and other reagents. (ii) Overview of the cell culture process in the microfluidic device: (Step 1) Bi-layer chip is fabricated with PDMS containing several channels on top of a glass slide. (Step 2) A mixture of cells and hydrogel precursor is fed from the cell inlet channel. (Step 3) A growth medium is supplied from the medium inlet channel for cell culture.

Mentions: Conventional macroscale 3D cell culture systems are not well suited to rapidly investigate the complex in vivo-like 3D microenvironments due to aforementioned limitations in imaging and HTS capability [19,76,79]. On the other hand, miniaturized 3D cell models significantly reduce assay volume and reagent consumption and provide excellent control over cellular microenvironments by precisely managing cell culture conditions in a combinatorial fashion within small dimensions. Due to the reduced sample volume required, we may be able to use expensive and scarce patient-derived cells, which may lead to enhanced predictability of in vivo drug responses in individuals [19]. Miniaturization of 3D cell models can reduce the time required for image acquisition and analysis, which makes 3D cell culture system more amenable to high-throughput HCI. Image acquisition would be much simpler due to the thin depth of Z-focus position of samples, leading to significantly increased signal-to-noise ratio [80]. Current miniaturized 3D cell culture models can be broadly classified into three categories: microwell platforms, cellular microarrays, and microfluidic devices (Figure 2). The following section summarizes these miniaturized 3D cell culture platforms and highlights several HCI assays demonstrated on these platforms (Table 4).


High Content Imaging (HCI) on Miniaturized Three-Dimensional (3D) Cell Cultures.

Joshi P, Lee MY - Biosensors (Basel) (2015)

Miniaturized 3D cell culture systems for HCI. (A) Microwell platform (Adapted from Ref [90] with permission of The Royal Society of Chemistry). Overview of hydrogel microwell arrays fabrication process: (Step 1) A polydimethylsiloxane (PDMS) stamp containing an array of micropillars is cast on a silicon master. (Steps 2 and 3) Poly(ethylene glycol) (PEG) gel is cross-linked to contain complementary microwell array topography using the PDMS stamp as a template. (Step 4) Individual cells are trapped on the hydrogel surface after swelling and washing of the surface. (B) Cellular microarrayson a functionalized glass slide. A mixture of cells and hydrogel precursor is printed on a glass slide coated with poly(styrene-co-maleic anhydride) (PS-MA). Various polymer coating is done on top of the PS-MA coating to attach different hydrogels to the glass slide. Cells are encapsulated in a hydrogel matrix, forming 3D structures after gelation (which occurs via various mechanisms). (C) Cellular microarrays on a micropillar/microwell chip platform (Adapted by permission from Macmillan Publishers Ltd: Nature Communications, Ref [84]). Cells mixed with hydrogel are printed on top of the micropillar chip. After gelation, the micropillar chip containing cells encapsulated in hydrogel is sandwiched with a complementary microwell chip containing growth media or other reagents. (D) Microfluidicdevice. (i) Top view of a bilayer microfluidic chip fabricated with PDMS on top of a glass slide. Several inlet and outlet channels provide parallel access to cell suspension, growth medium and other reagents. (ii) Overview of the cell culture process in the microfluidic device: (Step 1) Bi-layer chip is fabricated with PDMS containing several channels on top of a glass slide. (Step 2) A mixture of cells and hydrogel precursor is fed from the cell inlet channel. (Step 3) A growth medium is supplied from the medium inlet channel for cell culture.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4697144&req=5

biosensors-05-00768-f002: Miniaturized 3D cell culture systems for HCI. (A) Microwell platform (Adapted from Ref [90] with permission of The Royal Society of Chemistry). Overview of hydrogel microwell arrays fabrication process: (Step 1) A polydimethylsiloxane (PDMS) stamp containing an array of micropillars is cast on a silicon master. (Steps 2 and 3) Poly(ethylene glycol) (PEG) gel is cross-linked to contain complementary microwell array topography using the PDMS stamp as a template. (Step 4) Individual cells are trapped on the hydrogel surface after swelling and washing of the surface. (B) Cellular microarrayson a functionalized glass slide. A mixture of cells and hydrogel precursor is printed on a glass slide coated with poly(styrene-co-maleic anhydride) (PS-MA). Various polymer coating is done on top of the PS-MA coating to attach different hydrogels to the glass slide. Cells are encapsulated in a hydrogel matrix, forming 3D structures after gelation (which occurs via various mechanisms). (C) Cellular microarrays on a micropillar/microwell chip platform (Adapted by permission from Macmillan Publishers Ltd: Nature Communications, Ref [84]). Cells mixed with hydrogel are printed on top of the micropillar chip. After gelation, the micropillar chip containing cells encapsulated in hydrogel is sandwiched with a complementary microwell chip containing growth media or other reagents. (D) Microfluidicdevice. (i) Top view of a bilayer microfluidic chip fabricated with PDMS on top of a glass slide. Several inlet and outlet channels provide parallel access to cell suspension, growth medium and other reagents. (ii) Overview of the cell culture process in the microfluidic device: (Step 1) Bi-layer chip is fabricated with PDMS containing several channels on top of a glass slide. (Step 2) A mixture of cells and hydrogel precursor is fed from the cell inlet channel. (Step 3) A growth medium is supplied from the medium inlet channel for cell culture.
Mentions: Conventional macroscale 3D cell culture systems are not well suited to rapidly investigate the complex in vivo-like 3D microenvironments due to aforementioned limitations in imaging and HTS capability [19,76,79]. On the other hand, miniaturized 3D cell models significantly reduce assay volume and reagent consumption and provide excellent control over cellular microenvironments by precisely managing cell culture conditions in a combinatorial fashion within small dimensions. Due to the reduced sample volume required, we may be able to use expensive and scarce patient-derived cells, which may lead to enhanced predictability of in vivo drug responses in individuals [19]. Miniaturization of 3D cell models can reduce the time required for image acquisition and analysis, which makes 3D cell culture system more amenable to high-throughput HCI. Image acquisition would be much simpler due to the thin depth of Z-focus position of samples, leading to significantly increased signal-to-noise ratio [80]. Current miniaturized 3D cell culture models can be broadly classified into three categories: microwell platforms, cellular microarrays, and microfluidic devices (Figure 2). The following section summarizes these miniaturized 3D cell culture platforms and highlights several HCI assays demonstrated on these platforms (Table 4).

Bottom Line: High content imaging (HCI) is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates.Conventional HCI assays have been carried out on two-dimensional (2D) cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D) cell cultures.Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS).

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical & Biomedical Engineering, Cleveland State University, 1960 East 24th Street Cleveland, Ohio, OH 44115-2214, USA. p.joshi18@vikes.csuohio.edu.

ABSTRACT
High content imaging (HCI) is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates. Conventional HCI assays have been carried out on two-dimensional (2D) cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D) cell cultures. Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS). One attempt of making 3D cell culture amenable for HTS is to utilize miniaturized cell culture platforms. This review aims to highlight miniaturized 3D cell culture platforms compatible with current HCI technology.

Show MeSH
Related in: MedlinePlus