Microscale tissue engineering using gravity-enforced cell assembly

https://doi.org/10.1016/j.tibtech.2004.02.002Get rights and content

Abstract

Designing artificial microtissues by reaggregation of monodispersed primary cells, neoplastic or engineered cell lines is providing insight into cell–cell interactions and underlying regulatory networks. Recent advances in microtissue production have highlighted the potential of scaffold-free cell aggregates in maintaining tissue-specific functionality, supporting seamless integration of implants into host tissues, and providing complex feeder structures for difficult-to-differentiate cell types. Furthermore, these tissues are amenable to therapeutic and phenotype-modulating interventions using latest-generation transduction technologies. Microtissues produce therapeutic transgenes at increased levels and offer tissue-like assay environments to improve drug-function correlations in current discovery programs. Here, we outline scaffold-free microtissue design in liver, heart and cartilage, and discuss how this technology could significantly impact regenerative medicine.

Section snippets

Scaffold-free microtissues – hanging-drop technology revisited

Strategies using natural reaggregation potential to assemble monodispersed cells in a tissue-mimicking way represent a valuable extension of current scaffold-based tissue engineering initiatives. Scaffold-free reaggregation of cells to microtissues can occur following: (i) cultivation in shake flasks, gyratory shakers and roller bottles 13, 14, 15 or on non-adhesive surfaces [16]; (ii) centrifugation-based compression [17]; (iii) maintenance in cell-culture inserts [18]; or (iv)

Design of artificial hepatic tissues

The healthy liver is able to regenerate after injury. However, once damaged by fibrosis and cirrhosis – the result of a variety of chronic conditions, including alcohol abuse or infection with hepatitis virus B or C – the regeneration capacity of the liver is compromised. Liver transplantation is a routine treatment for end-stage liver disease, yet donor organ shortage continues to be a serious problem [26]. The liver has many crucial functions, including the production of clotting factors and

Artificial myocardial microtissue

Heart diseases such as myocardial infarction and heart failure are the most prevalent pathologies in industrialized countries. Loss of cardiomyocytes accounts for decreased myocardial function, which can result in total organ failure or trigger compensatory mechanisms such as hypertrophy of the remaining myocardium, activation of neurohumoral systems and/or autokrine or parakrine stimulation by various growth factors or cytokines [33]. The ultimate treatment of end-stage heart failure is heart

Microcartilage

Osteoarthritis and rheumatoid arthritis – the most prevalent disorders of the musculoskeletal system – result from the disturbance of tissue homeostasis in articular joints, and are diagnosed by joint pain, tenderness, movement limitations, as well as effusion and variable degrees of inflammation. Rheumatoid arthritis is characterized by chondrocytes that produce inflammatory signals and matrix metalloproteinases, which result in thinning of the collagen network, decrease of proteoglycan

Beyond tissue engineering: the future of microtissues in biopharmaceutical manufacturing and high-throughput drug discovery

Monodispersed cells growing in suspension and protein-free media are currently the gold standard for large-scale manufacturing of protein therapeutics [45]. Although key biopharmaceutical manufacturing parameters such as growth rate, cell density and specific productivity can be optimized by advanced bioprocess control or specific molecular interventions in production cell lines, the question remains as to whether specific productivity of suspension cells typically reached in classical

Concluding remarks

‘We are a self-assembling organism. That information is there to be captured and used.’ These were the words of William Haseltine, Chairman and CEO of Human Genome Sciences (http://www.hgsi.com/), who was among the first to coin the term regenerative medicine to describe new ways of teaching the body to heal itself [52]. Microtissue design consisting of reaggregation of biopsy-derived (stem) cell populations or (stem) cell lines harnesses the self-assembling programs of the organism to provide

Acknowledgements

We thank Lars K. Nielsen for providing the micrographs in Figure 3. We also thank David Fluri, Beat P. Kramer and Shizuka Hartenbach for critical comments on the manuscript. Work in the laboratory of M.F. is supported by the Swiss National Science Foundation (Grant No. 631065946).

References (70)

  • G.M. Keller

    In vitro differentiation of embryonic stem cells

    Curr. Opin. Cell Biol.

    (1995)
  • S. Petit-Zeman

    Regenerative medicine

    Nat. Biotechnol.

    (2001)
  • A. Bouche

    Xenotransplantation success for Immerge

    Nat. Biotechnol.

    (2002)
  • M.P. Lutolf

    Repair of bone defects using synthetic mimetics of collagenous extracellular matrices

    Nat. Biotechnol.

    (2003)
  • M. Fussenegger

    Controlled proliferation by multigene metabolic engineering enhances the productivity of Chinese hamster ovary cells

    Nat. Biotechnol.

    (1998)
  • S. Rafii et al.

    Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration

    Nat. Med.

    (2003)
  • A.A. Mangi

    Mesenchymal stem cells modified with Akt prevent remodeling and restore performance of infarcted hearts

    Nat. Med.

    (2003)
  • A. Abbott

    Cell culture: biology's new dimension

    Nature

    (2003)
  • C. Zandonella

    Tissue engineering: the beat goes on

    Nature

    (2003)
  • S.S. Kim

    Survival and function of hepatocytes on a novel three-dimensional synthetic biodegradable polymer scaffold with an intrinsic network of channels

    Ann. Surg.

    (1998)
  • R.J. Lee

    VEGF gene delivery to myocardium: deleterious effects of unregulated expression

    Circulation

    (2000)
  • T.P. Richardson

    Polymeric system for dual growth factor delivery

    Nat. Biotechnol.

    (2001)
  • E.R. Ochoa et al.

    An overview of the pathology and approaches to tissue engineering

    Ann. New York Acad. Sci.

    (2002)
  • Kelm, J.M. et al. Design of artificial myocardial microtissues. Tissue Eng. (in...
  • J.M. Kelm

    Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types

    Biotechnol. Bioeng.

    (2003)
  • K.S. Furukawa

    Formation of human fibroblast aggregates (spheroids) by rotational culture

    Cell Transplant.

    (2001)
  • S. Kale

    Three-dimensional cellular development is essential for ex vivo formation of human bone

    Nat. Biotechnol.

    (2000)
  • A. Muraglia

    Formation of a chondro-osseous rudiment in micromass cultures of human bone-marrow stromal cells

    J. Cell Sci.

    (2003)
  • S.B. Watzka

    Selection of viable cardiomyocytes for cell transplantation using three-dimensional tissue culture

    Transplantation

    (2000)
  • A.M. Wobus

    Embryonic stem cells and nuclear transfer strategies. Present state and future prospects

    Cells Tissues Organs

    (2000)
  • R. Bjerkvig

    Tumor cell invasion and angiogenesis in the central nervous system

    Curr. Opin. Oncol.

    (1997)
  • J. Itskovitz-Eldor

    Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers

    Mol. Med.

    (2000)
  • J.M. Kelm

    Synergies of microtissue design, viral transduction and adjustable transgene expression for regenerative medicine

    Biotechnol. Appl. Biochem.

    (2004)
  • D.L. Clarke

    Generalized potential of adult neural stem cells

    Science

    (2000)
  • T.R. Brazelton

    From marrow to brain: expression of neuronal phenotypes in adult mice

    Science

    (2000)
  • Cited by (0)

    View full text