Abstract: Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.
Abstract: Numerous investigations suggest that Mesenchymal
Stem Cells (MSCs) in general represent a valuable tool for therapy of
symptoms related to chronic inflammatory diseases. Blue Horizon
Stem Cell Therapy Program is a leading provider of adult and
children’s stem cell therapies. Uniquely we have safely and
efficiently treated more than 600 patients with documenting each
procedure. The purpose of our study is primarily to monitor the
immune response in order to validate the safety of intravenous
infusion of human umbilical cord blood derived MSCs (UC-MSCs),
and secondly, to evaluate effects on biomarkers associated with
chronic inflammation. Nine patients were treated for conditions
associated with chronic inflammation and for the purpose of antiaging.
They have been given one intravenous infusion of UCMSCs.
Our study of blood test markers of 9 patients with chronic
inflammation before and within three months after MSCs treatment
demonstrates that there is no significant changes and MSCs treatment
was safe for the patients. Analysis of different indicators of chronic
inflammation and aging included in initial, 24-hours, two weeks and
three months protocols showed that stem cell treatment was safe for
the patients; there were no adverse reactions. Moreover data from
follow up protocols demonstrates significant improvement in energy
level, hair, nails growth and skin conditions. Intravenously
administered UC-MSCs were safe and effective in the improvement
of symptoms related to chronic inflammation. Further close
monitoring and inclusion of more patients are necessary to fully
characterize the advantages of UC-MSCs application in treatment of
symptoms related to chronic inflammation.