Novel 3D Culture Technologies Utilizing Patient-Derived Tissue

July 12, 2016
Julia Kirshner, Ph.D. 

Human cells have been grown in culture since the 1950s. However, conventional cell culture models lack tissue microenvironment, and thus, have limited physiological relevance. Emerging 3D culture systems are starting to incorporate various components of tissue microenvironment to better replicate the biology of human tissues. zPREDICTA’s reconstructed organ platform mimics the native architecture of human tissues in an organ- and disease-specific manner, and thus, demonstrates high correlation with clinical response. This 45 minute educational webinar will summarize the current state of 3D culture technologies and will highlight relevant applications of each platform.

Q&A

What readout is compatible with reconstructed organ technology?
Reconstructed organ is compatible with any standard readout techniques, including but not limited to, flow cytometry, microscopy (brightfield, fluorescence, confocal), immunohistochemistry, nucleic acid analysis (RNAseq, qPCR, arrays, etc.), proteomics, cell-based assays (i.e. viability, proliferation, migration, etc.), and in vivo studies.

Cells can be isolated from reconstructed organ using non-enzymatic isolation solution that is compatible with flow cytometry and any other readout strategies that require intact surface receptors. Moreover, cells remain viable after isolation and can be subsequently used in both in vitro and in vivo studies.

Do primary, patient-derived cells maintain their phenotype in reconstructed organ cultures?
Yes. The proportion of each population is retained over the course of the culture (i.e. each cell population present in the patient material is maintained over the course of the culture in the same proportions as present in the starting material).

How long can cells remain viable in reconstructed organ cultures?
Viability of primary, patient-derived cells can be maintained for at least 28 days in reconstructed organ cultures.

What type of therapeutic agents can be tested in reconstructed organ?
Any type of therapeutic agent can be used in reconstructed organ: small molecules, antibodies, antibody-drug conjugates, biologic agents, cell therapeutics (i.e. CAR-T cells).

What is the thickness of reconstructed cultures?
The average thickness of the ECM is 800nm-1mm, which can be modulated depending on the study goals.

Is it possible to do high-resolution imaging and immunohistochemistry (IHC)?
Yes, in-matrix imaging is possible, which retains its native architecture, allowing analysis of cell-cell interactions and analysis of spatial orientation of cells and interactions between different cell types. Immunohistochemistry (IHC) is also possible.

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