Statistical Modeling of Single Target Cell Encapsulation

Moon, Sangjun; Ceyhan, Elvan; Gürkan, Umut A.; Demirci, Utkan

doi:10.1371/journal.pone.0021580

Cited by 49 publications

(49 citation statements)

References 35 publications

(65 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This result indicated that bioprinting did not significantly affect cell survival, which is consistent with our earlier studies. 9,33,51,52 Cells displayed typical healthy morphology generally observed in hydrogels (Figure 3B).…”

Section: Resultsmentioning

confidence: 81%

Engineering Anisotropic Biomimetic Fibrocartilage Microenvironment by Bioprinting Mesenchymal Stem Cells in Nanoliter Gel Droplets

et al. 2014

Self Cite

View full text Add to dashboard Cite

Over the past decade, bioprinting has emerged as a promising patterning strategy to organize cells and extracellular components both in two and three dimensions (2D and 3D) to engineer functional tissue mimicking constructs. So far, tissue printing has neither been used for 3D patterning of mesenchymal stem cells (MSCs) in multiphase growth factor embedded 3D hydrogels nor been investigated phenotypically in terms of simultaneous differentiation into different cell types within the same micropatterned 3D tissue constructs. Accordingly, we demonstrated a biochemical gradient by bioprinting nanoliter droplets encapsulating human MSCs, bone morphogenetic protein 2 (BMP-2), and transforming growth factor β1 (TGF- β1), engineering an anisotropic biomimetic fibrocartilage microenvironment. Assessment of the model tissue construct displayed multiphasic anisotropy of the incorporated biochemical factors after patterning. Quantitative real time polymerase chain reaction (qRT-PCR) results suggested genomic expression patterns leading to simultaneous differentiation of MSC populations into osteogenic and chondrogenic phenotype within the multiphasic construct, evidenced by upregulation of osteogenesis and condrogenesis related genes during in vitro culture. Comprehensive phenotypic network and pathway analysis results, which were based on genomic expression data, indicated activation of differentiation related mechanisms, via signaling pathways, including TGF, BMP, and vascular endothelial growth factor.

show abstract

Section: Resultsmentioning

confidence: 81%

Engineering Anisotropic Biomimetic Fibrocartilage Microenvironment by Bioprinting Mesenchymal Stem Cells in Nanoliter Gel Droplets

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…7 and Table 2). According to statistical modeling of cell encapsulation 94 and sorting 52 , the enrichment in heterogeneous cell mixtures (composed of two or more cell types) is limited by the coencapsulation probability of positive and negative cells. During the sorting step, droplets having at least one positive cell will be sorted regardless of the number of negative cells coencapsulated into the same droplet.…”

Section: Anticipated Resultsmentioning

confidence: 99%

Single-cell analysis and sorting using droplet-based microfluidics

et al. 2013

View full text Add to dashboard Cite

We present a droplet-based microfluidics protocol for high-throughput analysis and sorting of single cells. compartmentalization of single cells in droplets enables the analysis of proteins released from or secreted by cells, thereby overcoming one of the major limitations of traditional flow cytometry and fluorescence-activated cell sorting. as an example of this approach, we detail a binding assay for detecting antibodies secreted from single mouse hybridoma cells. secreted antibodies are detected after only 15 min by co-compartmentalizing single mouse hybridoma cells, a fluorescent probe and single beads coated with anti-mouse IgG antibodies in 50-pl droplets. the beads capture the secreted antibodies and, when the captured antibodies bind to the probe, the fluorescence becomes localized on the beads, generating a clearly distinguishable fluorescence signal that enables droplet sorting at ~200 Hz as well as cell enrichment. the microfluidic system described is easily adapted for screening other intracellular, cell-surface or secreted proteins and for quantifying catalytic or regulatory activities. In order to screen ~1 million cells, the microfluidic operations require 2–6 h; the entire process, including preparation of microfluidic devices and mammalian cells, requires 5–7 d.

show abstract

“…Moreover, a droplet has advantages to control and maintain cells within it, which allows effective handling for further processes such as separation and reactions. Encapsulation of a desired number of cells in a droplet is mainly described as a random process following Poisson’s distribution2. Although efficient ordination techniques for cell encapsulation have been reported in the literature, subsequent reactions are often needed to isolate droplets of interest345678.…”

mentioning

confidence: 99%

An on-chip imaging droplet-sorting system: a real-time shape recognition method to screen target cells in droplets with single cell resolution

Girault¹,

Kim²,

Arakawa³

et al. 2017

Sci Rep

View full text Add to dashboard Cite

A microfluidic on-chip imaging cell sorter has several advantages over conventional cell sorting methods, especially to identify cells with complex morphologies such as clusters. One of the remaining problems is how to efficiently discriminate targets at the species level without labelling. Hence, we developed a label-free microfluidic droplet-sorting system based on image recognition of cells in droplets. To test the applicability of this method, a mixture of two plankton species with different morphologies (Dunaliella tertiolecta and Phaeodactylum tricornutum) were successfully identified and discriminated at a rate of 10 Hz. We also examined the ability to detect the number of objects encapsulated in a droplet. Single cell droplets sorted into collection channels showed 91 ± 4.5% and 90 ± 3.8% accuracy for D. tertiolecta and P. tricornutum, respectively. Because we used image recognition to confirm single cell droplets, we achieved highly accurate single cell sorting. The results indicate that the integrated method of droplet imaging cell sorting can provide a complementary sorting approach capable of isolating single target cells from a mixture of cells with high accuracy without any staining.

show abstract

Statistical Modeling of Single Target Cell Encapsulation

Cited by 49 publications

References 35 publications

Engineering Anisotropic Biomimetic Fibrocartilage Microenvironment by Bioprinting Mesenchymal Stem Cells in Nanoliter Gel Droplets

Engineering Anisotropic Biomimetic Fibrocartilage Microenvironment by Bioprinting Mesenchymal Stem Cells in Nanoliter Gel Droplets

Single-cell analysis and sorting using droplet-based microfluidics

An on-chip imaging droplet-sorting system: a real-time shape recognition method to screen target cells in droplets with single cell resolution

Contact Info

Product

Resources

About