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Droplet size influences division of mammalian cell factories in droplet microfluidic cultivation
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.ORCID-id: 0000-0003-3618-9944
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.ORCID-id: 0000-0001-5232-0805
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi.
2016 (Engelska)Ingår i: Electrophoresis, ISSN 0173-0835, E-ISSN 1522-2683Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The potential of using droplet microfluidics for screening mammalian cell factories has been limited by the difficulty in achieving continuous cell division during cultivation in droplets. Here, we report the influence of droplet size on mammalian cell division and viability during cultivation in droplets. Chinese Hamster Ovary (CHO) cells, the most widely used mammalian host cells for biopharmaceuticals production were encapsulated and cultivated in 33, 180 and 320 pL droplets for 3 days. Periodic monitoring of the droplets during incubation showed that the cell divisions in 33 pL droplets stopped after 24 h, whereas continuous cell division was observed in 180 and 320 pL droplets for 72 h. The viability of the cells cultivated in the 33 pL droplets also dropped to about 50% in 72 h. In contrast, the viability of the cells in the larger droplets was above 90% even after 72 h of cultivation, making them a more suitable droplet size for 72-h cultivation. This study shows a direct correlation of microfluidic droplet size to the division and viability of mammalian cells. This highlights the importance of selecting suitable droplet size for mammalian cell factory screening assays.

Ort, förlag, år, upplaga, sidor
Wiley-VCH Verlagsgesellschaft, 2016.
Nationell ämneskategori
Bioprocessteknik
Forskningsämne
Bioteknologi
Identifikatorer
URN: urn:nbn:se:kth:diva-192574DOI: 10.1002/elps.201600316ISI: 000393298200009Scopus ID: 2-s2.0-84987648085OAI: oai:DiVA.org:kth-192574DiVA, id: diva2:971033
Anmärkning

QC 20160921

Tillgänglig från: 2016-09-15 Skapad: 2016-09-15 Senast uppdaterad: 2019-10-14Bibliografiskt granskad
Ingår i avhandling
1. Droplet microfluidics for single cell and nucleic acid analysis
Öppna denna publikation i ny flik eller fönster >>Droplet microfluidics for single cell and nucleic acid analysis
2016 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Droplet microfluidics is an emerging technology for analysis of single cells and biomolecules at high throughput. The controlled encapsulation of particles along with the surrounding microenvironment in discrete droplets, which acts as miniaturized reaction vessels, allows millions of particles to be screened in parallel. By utilizing the unit operations developed to generate, manipulate and analyze droplets, this technology platform has been used to miniaturize a wide range of complex biological assays including, but not limited to, directed evolution, rare cell detection, single cell transcriptomics, rare mutation detection and drug screening.

The aim of this thesis is to develop droplet microfluidics based methods for analysis of single cells and nucleic acids. In Paper I, a method for time-series analysis of mammalian cells, using automated fluorescence microscopy and image analysis technique is presented. The cell-containing droplets were trapped on-chip and imaged continuously to assess the viability of hundreds of isolated individual cells over time. This method can be used for studying the dynamic behavior of cells. In Paper II, the influence of droplet size on cell division and viability of mammalian cell factories during cultivation in droplets is presented. The ability to achieve continuous cell division in droplets will enable development of mammalian cell factory screening assays in droplets. In Paper III, a workflow for detecting the outcome of droplet PCR assay using fluorescently color-coded beads is presented. This workflow was used to detect the presence of DNA biomarkers associated with poultry pathogens in a sample. The use of color-coded detection beads will help to improve the scalability of the detection panel, to detect multiple targets in a sample. In Paper IV, a novel unit operation for label-free enrichment of particles in droplets using acoustophoresis is presented. This technique will be useful for developing droplet-based assays that require label-free enrichment of cells/particles and removal of droplet content. In general, droplet microfluidics has proven to be a versatile tool for biological analysis. In the years to come, droplet microfluidics could potentially be used to improve clinical diagnostics and bio-based production processes.

Ort, förlag, år, upplaga, sidor
Stockholm: KTH Royal Institute of Technology, 2016. s. 58
Serie
TRITA-BIO-Report, ISSN 1654-2312 ; 2016:17
Nyckelord
Acoustophoresis, Biomarker detection, Cell behavior analysis, Cell factories, Droplet microfluidics, Droplet PCR, High throughput biology, Label-free enrichment, Single cell analysis
Nationell ämneskategori
Övrig annan teknik
Forskningsämne
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-192668 (URN)978-91-7729-125-1 (ISBN)
Disputation
2016-10-21, Gard-aulan, Nobels väg 18, Solna, 10:00 (Engelska)
Opponent
Handledare
Anmärkning

QC 20160926

Tillgänglig från: 2016-09-26 Skapad: 2016-09-19 Senast uppdaterad: 2016-09-26Bibliografiskt granskad

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Periyannan Rajeswari, Prem Kumar

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Periyannan Rajeswari, Prem KumarJönsson, Håkan NAndersson Svahn, Helene
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Electrophoresis
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