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Living biointerfaces based on non-pathogenic bacteria support stem cell differentiation.

Jake J Hay1, Aleixandre Rodrigo-Navarro1, Karoliina Hassi1

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Genetically engineered bacteria, Lactococcus lactis, form biofilms expressing human fibronectin. These living interfaces support long-term human mesenchymal stem cell culture and osteoblastic differentiation, offering a novel biomaterial for regenerative medicine.

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Area of Science:

  • Biotechnology
  • Biomaterials Science
  • Stem Cell Biology

Background:

  • Mammalian cell culture requires biocompatible substrates.
  • Fibronectin is crucial for cell adhesion and differentiation.
  • Developing dynamic, long-term cell culture interfaces is challenging.

Purpose of the Study:

  • To engineer Lactococcus lactis to express human fibronectin for cell culture.
  • To evaluate the engineered bacteria as a substrate for human mesenchymal stem cell (hMSC) culture.
  • To assess the potential of this living interface for stem cell differentiation.

Main Methods:

  • Genetic engineering of Lactococcus lactis to express the III7-10 fibronectin fragment.
  • Biofilm formation on glass and synthetic polymers.
  • Long-term culture of hMSCs on engineered biofilms (up to 28 days).
  • Assessment of hMSC adhesion and osteoblastic differentiation (induced by BMP-2).

Main Results:

  • Engineered L. lactis formed stable, viable biofilms.
  • Biofilms supported long-term hMSC culture and adhesion.
  • The fibronectin-expressing biofilm was functionally equivalent to fibronectin-coated surfaces for osteoblastic differentiation.
  • The system demonstrated stability for up to 28 days.

Conclusions:

  • Engineered L. lactis biofilms provide a stable, living interface for hMSC culture.
  • This bacterial system effectively supports hMSC adhesion and osteoblastic differentiation.
  • The platform offers a dynamic substrate for stem cell applications and can be further engineered for controlled differentiation.