Engineered extracellular matrices with cleavable crosslinkers for cell expansion and easy cell recovery

Biomaterials. 2008 Dec;29(34):4521-31. doi: 10.1016/j.biomaterials.2008.08.008. Epub 2008 Sep 2.


An unmet need for expansion of primary cells and progenitor cells in three dimensions (3-D) is a synthetic mimic of the extracellular matrix (ECM) with user-controllable composition that would permit rapid recovery of viable cells under mild, non-enzymatic conditions. Three block copolymers based on disulfide-containing polyethylene glycol diacrylate crosslinkers were synthesized, and were used to crosslink thiol-modified hyaluronan and gelatin macromonomers in the presence of cells. The triblock PEGSSDA contained a single disulfide-containing block, the pentablock PEG(SS)(2)DA contained two disulfide blocks, and the heptablock PEG(SS)(3)DA contained three disulfide blocks. For each hydrogel composition, four cell types were encapsulated in 3-D, and growth and proliferation were evaluated. Murine NIH 3T3 fibroblasts, human HepG2 C3A hepatocytes, human bone marrow-derived mesenchymal stem cells (MSCs), and human umbilical vein endothelial cells (HUVECs) all showed excellent viability and growth during expansion in 3-D in the three disulfide block copolymer crosslinkers. After cell expansion, the hydrogels were dissociated using the thiol-disulfide exchange reaction in the presence of N-acetyl-cysteine or glutathione, which dissolved the hydrogel network. After dissolution, cells were recovered in high yield and with high viability by gentle centrifugation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biocompatible Materials / chemical synthesis*
  • Biocompatible Materials / chemistry
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Proliferation
  • Cell Survival
  • Cross-Linking Reagents*
  • Extracellular Matrix / chemistry*
  • Extracellular Matrix / metabolism*
  • Fibroblasts / cytology
  • Hepatocytes / cytology
  • Humans
  • Hydrogels / chemical synthesis
  • Hydrogels / chemistry
  • Mesenchymal Stem Cells / cytology
  • Mice
  • Polymers / chemical synthesis


  • Biocompatible Materials
  • Cross-Linking Reagents
  • Hydrogels
  • Polymers