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

Paper ID Volume ID Publish Year Pages File Format Full-Text
9660 641 2008 11 PDF Available
Title
Engineered extracellular matrices with cleavable crosslinkers for cell expansion and easy cell recovery
Abstract

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)2DA contained two disulfide blocks, and the heptablock PEG(SS)3DA 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.

Keywords
Disulfide-containing crosslinkers; Thiol–disulfide exchange reaction; Synthetic extracellular matrix; In situ crosslinking; Primary cells; Hyaluronic acid
First Page Preview
Engineered extracellular matrices with cleavable crosslinkers for cell expansion and easy cell recovery
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 29, Issue 34, December 2008, Pages 4521–4531
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering