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Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.ORCID iD: 0000-0003-0578-4003
2007 (English)In: BMC Biotechnology, ISSN 1472-6750, E-ISSN 1472-6750, Vol. 7, no 88Article in journal (Refereed) Published
Abstract [en]

Background: Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale.

Results: Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms.

Conclusion: Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.

Place, publisher, year, edition, pages
2007. Vol. 7, no 88
Keyword [en]
Cell analysis, Cell culture scaffold, Dipeptide hydrogel
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-10044DOI: 10.1186/1472-6750-7-88ISI: 000252971600001Scopus ID: 2-s2.0-39049149107OAI: oai:DiVA.org:kth-10044DiVA: diva2:202241
Note
QC 20100806Available from: 2009-03-09 Created: 2009-03-09 Last updated: 2017-12-13Bibliographically approved
In thesis
1. Photophysical and Chemical Approaches to Cellular Biophysics
Open this publication in new window or tab >>Photophysical and Chemical Approaches to Cellular Biophysics
2008 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

The central theme in this thesis is reversibility. Two main attempts has been made to approach reversibility in cellular systems from both chemical and physical points of view. Reversibility of immunolabeling of proteins on the cell surface has been adressed by development of new fluorescent substances optimized for CALI (Chromophore-Assisted Laser Inactivation of protein). Aluminum phthalocyanine (AlPc) is here identified to be a good candidate for a new generation of fluorophores for efficient hydroxyl radical generation. It is shown that cells can be reversibly labeled with antibody-AlPc conjugates. In experiments on living cells the AlPcs were not only active as classic fluorophores but also as photocatalytic substances with destaining properties. Reversibility of cell immobilization is also reported, where cells cultured in microstructures were immobilized and 3D supported using hydrogels. Hydrogel formulation and application was optimized to achieve a system where both viability and ease of use was satisfied. Gel reversibility was actualized with pH and enzyme treatment. The developped method offers the possibility of stop flow culturing cells in controlled and reusable 3D environments.

Place, publisher, year, edition, pages
Stockholm: KTH, 2008. v, 26 p.
Series
Trita-FYS, ISSN 0280-316X ; 2008:54
National Category
Biophysics
Identifiers
urn:nbn:se:kth:diva-10097 (URN)978-91-7412-200-5 (ISBN)
Presentation
2008-12-18, FA 32, AlbaNova Universitetscentrum, Roslagstullsbacken 21, Stockholm, 11:00 (English)
Opponent
Supervisors
Note
QC 20101102Available from: 2009-03-16 Created: 2009-03-16 Last updated: 2010-11-02Bibliographically approved
2. Studies of Cellular Responses to External Stimuli in Engineered Microenvironments
Open this publication in new window or tab >>Studies of Cellular Responses to External Stimuli in Engineered Microenvironments
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Stockholm: KTH, 2009. xvi, 59 p.
Series
Trita-FYS, ISSN 0280-316X ; 09:73
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-11819 (URN)978-91-7415-529-7 (ISBN)
Public defence
2010-01-15, Sal FD5, AlbaNova, Roslagstullsbacken 21, Stockholm, 13:00 (English)
Opponent
Supervisors
Note
QC 20100806Available from: 2009-12-29 Created: 2009-12-29 Last updated: 2010-08-06Bibliographically approved
3. Asymmetric Cellular Microenvironments
Open this publication in new window or tab >>Asymmetric Cellular Microenvironments
2008 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis presents methods to combine 3D cell culture, microfluidics and gradients on a controlled cellular scale. 3D cultures in biological extracellular matrix gels or synthetic gels bridge the gap between organ-tissue cultures and traditional 2D cultures. A device for embedding, anchoring and culturing cells in a controlled 3D flow through micro-environment was designed and evaluated. The device was realized using an etched silicon pillar flow chamber filled with gel mixed with cells. The pillars anchor and stabilize the gel as well as increase the surface to volume ratio, permitting higher surface flow rates and improving diffusion properties. Within the structure cells were still viable and proliferating after six days of cultivation, showing that it is possible to perform medium- to-long term cultivation of cells in a controlled 3D environment.

This concept was further developed to include controllable and time stable 3D microgradient environments. In this system stable diffusion gradients can be generated by the application of two parallel fluid flows with different composition against opposite sides of a gel plug with embedded cells. Culture for up to two weeks was performed showing cells still viable and proliferating. The cell tracer dye calcein was used to verify gradient formation as the fluorescent intensity in exposed cells was proportional to the position in the chamber. Cellular response to an applied stimulus was demonstrated by use of an adenosine triphosphate gradient where the onset of an intracellular calcium release also depends on cell position.

Place, publisher, year, edition, pages
Stockholm: KTH, 2008. vi, 33 p.
Series
Trita-FYS, ISSN 0280-316X ; 2007:87
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-4783 (URN)978-91-7178-848-1 (ISBN)
Presentation
(English)
Supervisors
Note
QC 20101119Available from: 2008-06-02 Created: 2008-06-02 Last updated: 2010-11-19Bibliographically approved

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