Much of the work conducted on adult stem cells has focused on mesenchymal stem cells (MSCs) found within the bone marrow stroma. Adipose tissue, like bone marrow, is derived from the embryonic mesenchyme and contains a stroma that is easily isolated. Preliminary studies have recently identified a putative stem cell population within the adipose stromal compartment. This cell population, termed processed lipoaspirate (PLA) cells, can be isolated from human lipoaspirates and, like MSCs, differentiate toward the osteogenic, adipogenic, myogenic, and chondrogenic lineages. To confirm whether adipose tissue contains stem cells, the PLA population and multiple clonal isolates were analyzed using several molecular and biochemical approaches. PLA cells expressed multiple CD marker antigens similar to those observed on MSCs. Mesodermal lineage induction of PLA cells and clones resulted in the expression of multiple lineage-specific genes and proteins. Furthermore, biochemical analysis also confirmed lineage-specific activity. In addition to mesodermal capacity, PLA cells and clones differentiated into putative neurogenic cells, exhibiting a neuronal-like morphology and expressing several proteins consistent with the neuronal phenotype. Finally, PLA cells exhibited unique characteristics distinct from those seen in MSCs, including differences in CD marker profile and gene expression.

/pdf/human-adipose-tissue-is-a-source-of-multipotent-stem-cells-18jo72s52v.pdf

Human Adipose Tissue Is a Source of Multipotent Stem Cells

We report here that cells co-purifying with mesenchymal stem cells--termed here multipotent adult progenitor cells or MAPCs--differentiate, at the single cell level, not only into mesenchymal cells, but also cells with visceral mesoderm, neuroectoderm and endoderm characteristics in vitro. When injected into an early blastocyst, single MAPCs contribute to most, if not all, somatic cell types. On transplantation into a non-irradiated host, MAPCs engraft and differentiate to the haematopoietic lineage, in addition to the epithelium of liver, lung and gut. Engraftment in the haematopoietic system as well as the gastrointestinal tract is increased when MAPCs are transplanted in a minimally irradiated host. As MAPCs proliferate extensively without obvious senescence or loss of differentiation potential, they may be an ideal cell source for therapy of inherited or degenerative diseases.

/pdf/pluripotency-of-mesenchymal-stem-cells-derived-from-adult-200s4qlizf.pdf

Pluripotency of mesenchymal stem cells derived from adult marrow

Cells undergo a variety of biological responses when placed in hypoxic conditions, including activation of signalling pathways that regulate proliferation, angiogenesis and death. Cancer cells have adapted these pathways, allowing tumours to survive and even grow under hypoxic conditions, and tumour hypoxia is associated with poor prognosis and resistance to radiation therapy. Many elements of the hypoxia-response pathway are therefore good candidates for therapeutic targeting.

Hypoxia — a key regulatory factor in tumour growth

Hydrolysis of inositol phospholipids by phospholipase C is initiated by either receptor stimulation or opening of Ca2+ channels. This was once thought to be the sole mechanism to produce the diacylglycerol that links extracellular signals to intracellular events through activation of protein kinase C. It is becoming clear that agonist-induced hydrolysis of other membrane phospholipids, particularly choline phospholipids, by phospholipase D and phospholipase A2 may also take part in cell signaling. The products of hydrolysis of these phospholipids may enhance and prolong the activation of protein kinase C. Such prolonged activation of protein kinase C is essential for long-term cellular responses such as cell proliferation and differentiation.

https://science.sciencemag.org/content/sci/258/5082/607.full.pdf

Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C

Adult marrow-derived Mesenchymal Stem Cells (MSCs) are capable of dividing and their progeny are further capable of differentiating into one of several mesenchymal phenotypes such as osteoblasts, chondrocytes, myocytes, marrow stromal cells, tendon-ligament fibroblasts, and adipocytes. In addition, these MSCs secrete a variety of cytokines and growth factors that have both paracrine and autocrine activities. These secreted bioactive factors suppress the local immune system, inhibit fibrosis (scar formation) and apoptosis, enhance angiogenesis, and stimulate mitosis and differentiation of tissue-intrinsic reparative or stem cells. These effects, which are referred to as trophic effects, are distinct from the direct differentiation of MSCs into repair tissue. Several studies which tested the use of MSCs in models of infarct (injured heart), stroke (brain), or meniscus regeneration models are reviewed within the context of MSC-mediated trophic effects in tissue repair.

/pdf/mesenchymal-stem-cells-as-trophic-mediators-1sjekxtzx0.pdf

Mesenchymal stem cells as trophic mediators.

Deletion Mutant of Protein Kinase Cpll Inhibits Insulin-Stimulated 2- Deoxyglucose Uptake in L6 Rat Skeletal Muscle Cells

Background: Adipose-derived stem cells (ADSC) were isolated and characterized from lean and obese subjects. We previously reported that distinct differences were observed in differentiating lean and obese preadipocytes. Protein kinase C delta (PKCδ) is alternatively spliced and has important roles in apoptosis. PKCδI promotes apoptosis and PKCδVIII promotes survival. Our previous data indicated an increase in the survival kinase, PKCδVIII in ADSC derived from an obese donor. We also determined that obese adipocytes were resistant to apoptosis. Here, we determine the relationship between a survival kinase PKCδVIII and hTERT expression in adipose derived stem cells from a lean and obese subject.
 Methods: We evaluated the telomerase activity and human telomerase reverse transcriptase (hTERT) expression in lean and obese ADSC. The lean and obese ADSC were purchased as cryopreserved cells from ZenBio™ (Research Triangle Park, NC, USA). Analyses were performed using PRISM™ software and analyzed using two-tailed Student’s t -test.
 Results: We observed an increase in telomerase in differentiating obese ADSC using western blot analysis. We determined the levels of hTERT splice variants. hTERT α+/β+ splice variant was increased after transfected of PKCδVIII. We next determined whether PKCδVIII over-expression affected the levels of telomerase. The results indicate an increase in telomerase with PKCδVIII over-expression.
 Conclusions: Over-expression of PKCδVIII in lean ADSC substantially increased expression of hTERT and telomerase. The decreased senescence seen in obese ADSC may in part be attributed to PKCδVIII. Obese ADSC undergo lower senescence and may have increased growth potential. These results propose a larger epigenetic modification in obese ADSC compared to lean ADSC.

Protein kinase C delta (PKCδ) splice variant modulates senescence via hTERT in adipose-derived stem cells

Ovarian cancer remains the most common cause of death from gynecologic malignancy in US. The impact of obesity on the incidence and treatment of gynecologic cancer is growing in importance. Little is known about the molecular factors obesity imparts to the cancer niche that account for this relationship. Adipose-derived stem cells (ADSC) can be isolated from stromal vascular fraction of abdominal white adipose tissue. We isolated and differentiated ADSC into adipocytes and previously showed that ADSC derived from lean and obese patients have distinct differences in gene expression. We evaluated the possibility that the secretome of obese ADSC could have paracrine effects on ovarian cells promoting growth and development of cancer cells. We show that NEAT1 levels secreted by ADSC from obese patients (ADSCo) were considerably higher than from normal ADSC both in cells and its secretome. Ovarian cancer high mortality rates are further exacerbated by cisplatin-resistant cancers. Our data shows cisplatin-resis...

PKCδVIII and Bcl2 Increase Ovarian Cancer Survival via LncRNA NEAT1 Secreted by Obese Adipose Derived Stem Cells

The protein kinase C (PKC)-βII modulates glucose-induced cell proliferation in A10 cells, a clonal cell line of vascular smooth muscle cells (VSMC) from rat aorta, which were studied by overexpressing PKC-βII and using a PKC-βII specific inhibitor (CG53353). PKC-βII overexpression was verified by the fourfold increase of PKC enzyme activity and PKC-βII immunoreactivity. Overexpression of PKC-βII attenuated A10 cell proliferation and DNA synthesis through the suppression of cell cycle progression inhibiting the entry of cells into the S phase. High glucose (25 mM) increased and accelerated cell proliferation, DNA synthesis, and the percentage of cells entering the S phase in A10. High glucose down-regulated PKC-βII in A10 cells during the first cell cycle after cell synchronization. CG53353 inhibited glucose-induced cell proliferation and DNA synthesis specifically. These results suggest that PKC-βII has inhibitory functions as a cell cycle checkpoint mediator during the late G1 phase and may regulate the ...

The Involvement of Protein Kinase C-βII in Glucose-Induced Rat Vascular Smooth Muscle Cell Proliferation

Provided herein are exosomal compositions and exsosomal lncRNA compositions and formulations thereof. Also provided herein are methods of treating a wound in a subject in need thereof that can contain the step of administering an exosomal composition and/or exsosomal lncRNA compositions or formulation thereof to a wound in a subject in need thereof.

Denise R. Cooper

Papers

Deletion Mutant of Protein Kinase Cpll Inhibits Insulin-Stimulated 2- Deoxyglucose Uptake in L6 Rat Skeletal Muscle Cells

Protein kinase C delta (PKCδ) splice variant modulates senescence via hTERT in adipose-derived stem cells

PKCδVIII and Bcl2 Increase Ovarian Cancer Survival via LncRNA NEAT1 Secreted by Obese Adipose Derived Stem Cells

The Involvement of Protein Kinase C-βII in Glucose-Induced Rat Vascular Smooth Muscle Cell Proliferation

Adipose derived stem cell exosomes and uses thereof