Cellular and Molecular Life Sciences

, Volume 73, Issue 7, pp 1479–1488 | Cite as

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

  • Siqin Wu
  • Jan Johansson
  • Outi Hovatta
  • Anna Rising
Original Article

Abstract

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.

Keywords

hESC iPSC Self-renewal Differentiation Enzymatic passaging Culture matrices 

Abbreviations

2D

Two-dimensional

3D

Three-dimensional

BAC

Bacterial artificial chromosome

BMP4

Bone morphogenetic protein 4

cTnT

Cardiac troponin T

FGF2

Basic fibroblast growth factor

FOXA2

Forkhead box A2

hPSC

Human pluripotent stem cell

hESC

Human embryonic stem cell

hiPSC

Human induced pluripotent stem cell

NKX2.5

NK2 homeobox 5

OCT4

Octamer-binding transcription factor 4

PAX6

Paired box 6

SOX17

SRY (sex determining region Y) Box 17

SSEA-1

Stage-specific embryonic antigen 1

SSEA-4

Stage-specific embryonic antigen 4

Supplementary material

18_2015_2053_MOESM1_ESM.docx (7.9 mb)
Supplementary material 1 (DOCX 8114 kb)

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Copyright information

© Springer Basel 2015

Authors and Affiliations

  • Siqin Wu
    • 1
  • Jan Johansson
    • 1
    • 2
    • 3
  • Outi Hovatta
    • 4
  • Anna Rising
    • 1
    • 2
  1. 1.Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society (NVS), Center for Alzheimer ResearchKarolinska InstitutetStockholmSweden
  2. 2.Department of Anatomy, Physiology and BiochemistrySwedish University of Agricultural SciencesUppsalaSweden
  3. 3.Institute of Mathematics and Natural SciencesTallinn UniversityTallinnEstonia
  4. 4.Division of Obstetrics and Gynecology, Department of Clinical Sciences, Intervention and TechnologyKarolinska Institutet and Karolinska University HospitalStockholmSweden

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