Researchers are exploring young blood as a potential fountain of youth. Studies have demonstrated its effect on improving remyelination, increasing brain plasticity, and stimulating neuron growth; yet its exact mechanism remains elusive.
Passegue and her team first attempted to rejuvenate aged hematopoietic stem cells in mice through exercise or caloric restriction, but were unsuccessful in doing so.
Human umbilical cord blood plasma (hUCBP)
Human umbilical cord blood plasma (hUCBP) is an abundant source of mononuclear cells and growth factors that have the ability to stimulate cell proliferation, slow age-related symptoms and boost immunity – among its many unique multifunctional benefits such as scavenging free radicals, encouraging cell repair/regeneration processes and strengthening immune function – making hUCBP an attractive candidate therapeutic treatment candidate for various diseases or injuries.
Recent studies have demonstrated that human umbilical cord blood mononuclear cells (hUCB-MNCs) can significantly extend mouse longevity. Furthermore, hUCB-MNCs promote regeneration of damaged tissue regeneration while speeding cell repair through several mechanisms including antioxidant effects, immunomodulation, promotion of angiogenesis, and secretion of trophic factors.
Ageing can cause stem cell proliferation to decrease and an accumulation of beta-amyloid plaques to build up, leading to Alzheimer’s disease. Current research is exploring whether cord blood stem cells could provide a therapy that treats this progressive illness; additionally they offer anti-inflammatory benefits that could delay its progress.
Scientists conducted a study wherein they injected old mice with either saline or human umbilical cord blood plasma from young adults. They observed that injecting old mice with plasma from young adults rejuvenated them significantly, improving performance on maze tests and rotarod tests. Researchers then analyzed proteins present in plasma samples from young adults; one named TIMP-2 proved responsible for this rejuvenation effect.
Plasma from human umbilical cord blood has been demonstrated to be an extremely potent trophic factor that helps promote the proliferation of mesenchymal stem cells, human dental stem cells and T-lymphocytes in vitro. With relatively low immunogenicity levels making this donor attractive for immunomodulatory transplantation; however it must first be fully matched with their recipient recipient before proceeding.
TIMP-2
Human TIMP-2 gene encodes a protein with a cysteine-rich domain that regulates hippocampal extracellular matrix (ECM) components involved in memory and plasticity processes. We evaluated its effects on ECM components among wild-type (WT) and TIMP2fl/fl mice exposed to chronic stressor. Furthermore, RNA sequencing was done to identify differentially expressed genes between WT and TIMP2fl/fl hippocampi, followed by GSEA analysis to ascertain whether their expression changes reflected altered biological processes or cell components.
We modified the codon sequence of wild type TIMP-2 cDNA to increase expression levels in both HEK-293-F and CHO-S cells using sulfobetaine peptide reagent, known for reducing cell clumping and increasing recombinant protein yield. As a result, codon-optimized rhTIMP-2-6XHis could be isolated with high yield and high purity from transiently transfected HEK-293-F cells as well as stable transfected CHO-S cells with high yield and good purity; additionally it showed outstanding MMP inhibitory activity when tested against Eurofins Panlabs MMP assay.
To evaluate recombinant proteins, powdered rhTIMP-2-6XHis was resuspended at equivalent concentrations in buffers of pH 4, 7, and 10 and subjected to CD spectral analysis to observe any change in secondary structure caused by pH-dependence; no such changes occurred, suggesting it can be safely used within physiological or charged formulation excipients and buffers.
Pretreatment with low nanomolar amounts of rhTIMP-2-6XHis, in low nanomolar quantities, inhibited both A549 lung cancer cells and JygMC(A) triple-negative breast cancer cells with low nanomolar amounts reduced their proliferation to basal (unstimulated) levels indicating its potent tumor suppressive properties that could potentially be used against multiple forms of cancers.
GM-CSF
GM-CSF is an immunomodulatory cytokine that promotes growth and differentiation of granulocytes and macrophages (Mph), as well as their antimicrobial, proinflammatory and microbicidal functions. Increased expression of costimulatory molecules in APCs may increase expression and induce phagocytosis by improving surface adhesion of neutrophils; production of nitric oxide and reactive oxygen species also increase; plus it promotes binding between neutrophils and endothelial cells is promoted as well. GM-CSF has been found to play an essential role in mediating inflammation associated with age and age-related diseases, while also contributing to inflammatory reactions against viral infections. Studies have demonstrated that infection with Hepatitis C Virus (HCV) impairs GM-CSFR signaling in monocyte derived dendritic cells, leading to reduced ability of these cells to secrete IL-12 and induce Th1 differentiation and phagocytosis. Human Cymegalovirus Virus (HCMV) infection also impairs ex vivo dendritic cell function due to secreted cytokines which block STAT5 phosphorylation thereby rendering this functionless.
Studies on mice have demonstrated that deficiency of GM-CSF leads to increased susceptibility for specific pulmonary infections and impaired hematopoiesis, as well as impaired immune responses against IAV by activating the NLRP3 inflammasome in AMph and thus creating an inflammatory feedback loop that further worsens inflammation and could even cause severe lung damage.
Regenerative effects of GM-CSF have been attributed to its ability to stimulate stromal cell proliferation. Stromal cells play an essential role in tissue repair and regeneration, producing nitric oxide that improves circulation and blood flow while preventing clot formation in the brain. Such benefits may help alleviate Alzheimer’s symptoms; clinical trials for treatment with GM-CSF are currently taking place to test this theory.
MANF
MANF is a neurotrophic factor that enhances differentiation and migration of neural progenitor cells (NPCs). Furthermore, it protects against oxygen-glucose deprivation-induced injury by modulating p38-MAPK signaling pathways. Furthermore, MANF improves browning of subcutaneous white adipose tissue under inguinal subcutaneous white fat tissue in mice as well as improves their insulin resistance.
MANF can also demonstrate its cytoprotective activity against ischemia by inhibiting cell necrosis/apoptosis in rat brain ischemic model, while upregulating it in neurons and glial cells after injury from stroke-induce stroke-induced rats. Recombinant MANF injection improved neurological deficits such as Bederson’s neurological score test scores as well as elevated body swing test (EBST) scores by improving upregulation of p38-MAPKs such as Akt and Erk proteins. This effect was mediated through upregulation of p38-MAPKs, Akt, and Erk.
In another study, it was demonstrated that MANF significantly facilitated behavioral deficit reversal in MPTP/MPP+-induced PD mice by increasing BDNF expression and activating Akt/mTOR and Erk/mTOR signaling pathways. Furthermore, it attenuated Ab toxicity while decreasing expression of cleaved caspase-3 expression and protecting against oxidative stress while also regulating UPR-related genes in neurons.
MANF is produced by melanotrophs of the hypothalamus and found in the pituitary gland, where it plays an essential role in sexual behavior, social recognition and stress response. MANF is also expressed in neurons of the PVN and supraoptic nuclei (SON) parts of the hypothalamus, where it plays an essential role. Furthermore, its physiological functions extend to various organs and tissues throughout the body such as the heart, liver, and pancreas. Circulating levels of MANF rise during fasting and correlate to increases in adiponectin levels, making MANF an essential regulator of human metabolism and aging. Furthermore, its CXXC motif has been shown to reduce endogenous ER stress.
Klotho
Klotho (a-Klotho), also referred to as an anti-aging molecule, regulates cell function and lifespan by producing protein hormones which are secreted into our systems and then broken down further as needed into hormones. Enhancing synaptic plasticity and cognitive performance as well as strengthening neural resistance against aging, Alzheimer’s Disease (AD), and Parkinson’s disease is another benefit of brain training. Klotho can increase GluN2B levels in neurons and the hippocampus, elevate N-methyl-d-aspartate receptor activity, which enhances long-term potentiation, enhance cellular uptake of calcium, phosphate, and vitamin D, activate the mTOR pathway and NF-B signaling as well as provide long-term potentiation benefits. Klotho can be found in kidney, choroid plexus and cerebrospinal fluid. As we age, its expression decreases due to diabetes, chronic kidney disease (CKD), neurodegeneration and other vascular diseases; its anti-aging benefits may also include its protective role against renal damage. Balance of mitochondrial structure and function, preservation of aqueous ion homeostasis, promotion of redox balance, and inhibition of oxidative stress are among its many beneficial impacts. Furthermore, calcium supplements may play a significant role in the regulation of calcification, intima hyperplasia and endothelial dysfunction. Klotho is an integral component of a healthy cardiovascular system and can be enhanced through various drugs including renin inhibitors, statins, mTOR inhibitors, GLP-1 agonists and PPAR-g agonists. Preclinical studies indicate that Klotho can also enhance protective antioxidant enzyme expression as well as release more nitric oxide from its reservoir.
Klotho also regulates autophagy, an essential cell-recycling mechanism that removes damaged organelles and proteins, protects against oxidative damage, and preserves biological functions. Klotho increases autophagy by activating AMPK pathway while inhibiting ERK signaling pathway; additionally it protects renal tubular cell damage through regulation of Akt and mTOR phosphorylation as well as increasing microglial activation in an Alzheimer’s model mouse called the APP/PS1 mouse model; additionally its upregulation has improved cognition through inhibiting NLRP3 encouraging microglial differentiation as well as controlling expression of an Ab transporter protein transporter protein.
