Efficient passage of human pluripotent stem cells on spider silk matrices under xeno-free conditions

Sammanfattning

Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine and pharmaceutical development. Such applications require cell culture methods and reagents that are chemically defined, xeno-free, scalable, and low-cost. Herein, we describe non-mechanical passaging of hPSCs on spider silk films under chemically defined and xeno-free conditions. The cells were dissociated into single cells or small aggregates using Accutase or enzyme-free dissociation buffer and then passaged to spider silk films, where they expanded in monolayers until they covered the surface. Cells cultured over 10 passages on spider silk film remained karyotypically normal and pluripotent. In conclusion, a novel method for passaging dissociated hPSCs under conditions that are compatible with clinical applications is presented. The method is cost-efficient and may be useful for both research and clinical applications.

Nyckelord

hESC; iPSC; Self-renewal; Differentiation; Enzymatic passaging; Culture matrices

Publicerad i

Cellular and Molecular Life Sciences
2016, Volym: 73, nummer: 7, sidor: 1479-1488
Utgivare: SPRINGER BASEL AG

SLU författare

• Johansson, Jan

  • Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
    
  • Karolinska institutet
  • Tallinna Ülikool

• Rising, Anna

  • Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
    
  • Karolinska institutet

UKÄ forskningsämne

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
Cell- och molekylärbiologi


Publikationens identifierare

DOI: https://doi.org/10.1007/s00018-015-2053-5

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/80754