موجز عن البحث:
From a perspective of manufacturer,
procurement of bone marrow aspirates for isolation of mesenchymal
stromal/stem cells (MSC) is challenging. The MSC isolated from adult donors
have lower proliferation capacity than the cells isolated form young donors of
pediatric age. To obtain more MSC from young healthy donors for allogeneic
therapy on multiple patients, umbilical cord (UC) seems to be the best
alternative. Here, we describe an easy, cost-effective and reproducible
protocol of isolation of the MSC from Wharton’s Jelly (WJ) in UC.
موجز عن البحث:
AIM: Umbilical cord contains, within Wharton’s
jelly (WJ), multipotent mesenchymal stromal/stem cells (MSCs) of fetal origin
that can be isolated and expanded in vitro with a minimal manipulation and
very high efficiency. Our aim was to develop a highly reproducible protocol
that has the unique potential to be scaled up and adapted to cGMP
requirements for the use in cellular therapy.
RESULTS: We found that derivation of WJ MSCs
under defined conditions in low oxygen resulted in several folds higher
populations of MSCA-1(+) cells (6.0-19.2%) when compared with WJ MSCs derived
in the presence of serum (0.1-2.8%) or clinical-grade bone marrow (BM) MSCs
cultured under atmospheric O2 (20%). We demonstrate that WJ MSCs derived
following our protocol display antiproliferative activity similar to
clinical-grade BM MSCs. We also show that these WJ MSCs can be differentiated
into adipo-, chondro- and osteo-genic lineages.
CONCLUSION: Easy accessibility, abundance and
genetic ‘naivety’ make WJ MSCs logistically a more attractive source for
clinical applications than BM MSCs.
موجز عن البحث:
The epigenetic background of
pluripotent stem cells can influence transcriptional and functional behavior.
Most of these data have been obtained in standard monolayer cell culture
systems. In this study, we used exome sequencing, array comparative genomic hybridization
(CGH), miRNA array, DNA methylation array, three-dimensional (3D) tissue
engineering, and immunostaining to conduct a comparative analysis of two
induced pluripotent stem cell (iPSC) lines used in engineering of 3D human
epidermal equivalent (HEE), which more closely approximates epidermis. Exome
sequencing and array CGH suggested that their genome was stable following 3
months of feeder-free culture. While the miRNAome was also not affected, ≈7%
of CpG sites were differently methylated between the two lines. Analysis of
the epidermal differentiation complex, a region on chromosome 1 that contains
multiple genes involved in skin barrier maturation (including trichohyalin,
TCHH), found that in one of the iPSC clones (iKCL004), TCHH retained a DNA
methylation signature characteristic of the original somatic cells, whereas
in other iPSC line (iKCL011), the TCHH methylation signature matched that of
the human embryonic stem cell line KCL034. The difference between the two
iPSC clones in TCHH methylation did not have an obvious effect on its
expression in 3D HEE, suggesting that differentiation and tissue formation
may mitigate variations in the iPSC methylome.