Abstract
In situ detection of MSCs remains difficult and warrants additional methods to aid with their characterization in vivo. Two-photon confocal laser scanning microscopy (TPM) and second harmonic generation (SHG) could fill this gap. Both techniques enable the detection of cells and extracellular structures, based on intrinsic properties of the specific tissue and intracellular molecules under optical irradiation. TPM imaging and SHG imaging have been used for label-free monitoring of stem cells differentiation, assessment of their behavior in biocompatible scaffolds, and even cell tracking in vivo. In this study, we show that TPM and SHG can accurately depict the umbilical cord architecture and visualize individual cells both in situ and during culture initiation, without the use of exogenously applied labels. In combination with nuclear DNA staining, we observed a variance in fluorescent intensity in the vessel walls. In addition, antibody staining showed differences in Oct4, αSMA, vimentin, and ALDH1A1 expression in situ, indicating functional differences among the umbilical cord cell populations. In future research, marker-free imaging can be of great added value to the current antigen-based staining methods for describing tissue structures and for the identification of progenitor cells in their tissue of origin.
Highlights
Stem cells originating from perinatal tissues such as the umbilical cord (UC) are being intensively studied for application in regenerative medicine
We show that Two-photon confocal laser scanning microscopy (TPM) and second harmonic generation (SHG) can accurately depict the umbilical cord architecture and visualize individual cells both in situ and during culture initiation, without the use of exogenously applied labels
We show that TPM is an elegant tool to characterize UC stem cells in situ, with the potential for parallel use with conventional imaging and staining techniques
Summary
Stem cells originating from perinatal tissues such as the umbilical cord (UC) are being intensively studied for application in regenerative medicine. The derivation of multipotent cells from the UC matrix or Wharton’s jelly (WJ) was first described about a decade ago by Mitchell et al and Romanov et al, reporting the isolation of stromal cells with a mesenchymal-like phenotype (WJ-MSCs) [9, 10]. Following their discovery, their potent preclinical potential as well as their superior culture properties over adult (bone-marrow-derived) mesenchymal stem cells (MSCs) has been extensively described [11,12,13,14,15,16]. In contrast to this extensive characterization in culture, the biology of WJ-MSCs in situ and their transition from tissue into culture remain poorly understood
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