A matter of identity — Phenotype and differentiation potential of human somatic stem cells

S.E.P New,C Alvarez-Gonzalez,B Vagaska,S.G Gomez,N.W Bulstrode,A Madrigal,P Ferretti

doi:10.1016/j.scr.2015.04.003

S.E.P New, C Alvarez-Gonzalez + Show 5 more

Open Access

https://doi.org/10.1016/j.scr.2015.04.003

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Journal: Stem Cell Research	Publication Date: Apr 25, 2015
Citations: 32	License type: cc-by

Affiliation: University College London

Abstract

Human somatic stem cells with neural differentiation potential can be valuable for developing cell-based therapies, including treatment of birth-related defects, while avoiding issues associated with cell reprogramming. Precisely defining the “identity” and differentiation potential of somatic stem cells from different sources, has proven difficult, given differences in sets of specific markers, protocols used and lack of side-by-side characterization of these cells in different studies. Therefore, we set to compare expression of mesenchymal and neural markers in human umbilical cord-derived mesenchymal stem cells (UC-MSCs), pediatric adipose-derived stem cells (p-ADSCs) in parallel with human neural stem cells (NSCs). We show that UC-MSCs at a basal level express mesenchymal and so-called “neural” markers, similar to that we previously reported for the p-ADSCs. All somatic stem cell populations studied, independently from tissue and patient of origin, displayed a remarkably similar expression of surface markers, with the main difference being the restricted expression of CD133 and CD34 to NSCs. Expression of certain surface and neural markers was affected by the expansion medium used. As predicted, UC-MSCs and p-ADSCs demonstrated tri-mesenchymal lineage differentiation potential, though p-ADSCs display superior chondrogenic differentiation capability. UC-MSCs and p-ADSCs responded also to neurogenic induction by up-regulating neuronal markers, but crucially they appeared morphologically immature when compared with differentiated NSCs. This highlights the need for further investigation into the use of these cells for neural therapies. Crucially, this study demonstrates the lack of simple means to distinguish between different cell types and the effect of culture conditions on their phenotype, and indicates that a more extensive set of markers should be used for somatic stem cell characterization, especially when developing therapeutic approaches.

Highlights

Extended Methods Isolation and culturing of neural stem cells (NSCs) All procedures involving human tissue were carried out in accordance to the Human Tissue Act 2006
The human neural stem cell lines lines were derived from brain and spinal cords from human embryos and fetuses between Carnegie stage (CS) 18 and CS22
After removing the supernatant and the floating mature adipocytes, cell pellets were incubated with red blood cell lysis buffer (Roche) at room temperature for 5 minutes and centrifuged again

Summary

Introduction

Extended Methods Isolation and culturing of neural stem cells (NSCs) All procedures involving human tissue were carried out in accordance to the Human Tissue Act 2006. The dissected tissues were minced and transferred into 0.5 ml accutase solution (Stem cell pro, Gibco, #A1110501) using a sterile Pasteur pipette and incubated at 37°C for 5-10 minutes with occasional trituration with a P1000 pipette to obtain a single cell suspension. This consisted RHB-A medium (Stem Cell Sciences Ltd, UK), supplemented with L-glutamine (2 mM final, Invitrogen), penicillin-streptomycin (10 mL/L final, Invitrogen), N2 (1X final, PAA), B27 (1X final, PAA), recombinant human FGF (20 ng/mL final, Peprotech), recombinant human EGF (20 ng/mL final Peprotech), Heparin Sulfate (5 μg/mL final, Sigma), Laminin L2020 (10 μg/mL final, Sigma) and BSA Fraction V (100 μg/mL final, Sigma).

Results

Conclusion