Hyaluronan in mesenchymal stromal cell lineage differentiation from human pluripotent stem cells: application in serum free culture

Paul A De Sousa*, Leo Perfect, Jinpei Ye, Kay Samuels, Ewa Piotrowska, Martin Gordon, Ryan Mate, Elsa Abranches, Thomas M Wishart, David H Dockrell, Aidan Courtney

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

BACKGROUND: Hyaluronan (HA) is an extracellular glycosaminoglycan polysaccharide with widespread roles throughout development and in healthy and neoplastic tissues. In pluripotent stem cell culture it can support both stem cell renewal and differentiation. However, responses to HA in culture are influenced by interaction with a range of cognate factors and receptors including components of blood serum supplements, which alter results. These may contribute to variation in cell batch production yield and phenotype as well as heighten the risks of adventitious pathogen transmission in the course of cell processing for therapeutic applications. MAIN: Here we characterise differentiation of a human embryo/pluripotent stem cell derived Mesenchymal Stromal Cell (hESC/PSC-MSC)-like cell population by culture on a planar surface coated with HA in serum-free media qualified for cell production for therapy. Resulting cells met minimum criteria of the International Society for Cellular Therapy for identification as MSC by expression of. CD90, CD73, CD105, and lack of expression for CD34, CD45, CD14 and HLA-II. They were positive for other MSC associated markers (i.e.CD166, CD56, CD44, HLA 1-A) whilst negative for others (e.g. CD271, CD71, CD146). In vitro co-culture assessment of MSC associated functionality confirmed support of growth of hematopoietic progenitors and inhibition of mitogen activated proliferation of lymphocytes from umbilical cord and adult peripheral blood mononuclear cells, respectively. Co-culture with immortalized THP-1 monocyte derived macrophages (Mɸ) concurrently stimulated with lipopolysaccharide as a pro-inflammatory stimulus, resulted in a dose dependent increase in pro-inflammatory IL6 but negligible effect on TNFα. To further investigate these functionalities, a bulk cell RNA sequence comparison with adult human bone marrow derived MSC and hESC substantiated a distinctive genetic signature more proximate to the former.

CONCLUSION: Cultivation of human pluripotent stem cells on a planar substrate of HA in serum-free culture media systems is sufficient to yield a distinctive developmental mesenchymal stromal cell lineage with potential to modify the function of haematopoietic lineages in therapeutic applications.

Original languageEnglish
Article number130
Pages (from-to)1-21
Number of pages21
JournalStem Cell Research and Therapy
Issue number1
Early online date3 May 2024
Publication statusE-pub ahead of print - 3 May 2024

Keywords / Materials (for Non-textual outputs)

  • Humans
  • Mesenchymal Stem Cells/metabolism
  • Hyaluronic Acid/pharmacology
  • Cell Differentiation
  • Pluripotent Stem Cells/metabolism
  • Culture Media, Serum-Free/pharmacology
  • Cell Lineage
  • Cells, Cultured
  • Cell Culture Techniques/methods
  • Coculture Techniques


Dive into the research topics of 'Hyaluronan in mesenchymal stromal cell lineage differentiation from human pluripotent stem cells: application in serum free culture'. Together they form a unique fingerprint.

Cite this