The application of pluripotent stem cells that received the most attention in recent years is as a novel source of cells for cell replacement therapy for the treatment of a wide range of debilitating diseases.
Type I diabetes, cardiovascular disease, Parkinson’s disease, blood cell diseases, and certain types of liver disease are considered candidates for cell replacement therapy. nEPS may be used to treat diseases of the nervous system, vascular, muscle and bone diseases, autoimmune system and also of the damaged organs.
Type I Diabetes
Diabetes mellitus has no permanent cure to date. One of the plausible therapeutic strategies to achieve this goal is through pancreatic stem cell transplantation. There are various stem cell sources and manipulation techniques that can be used to generate functional β-cells in a safe and efficient manner. And in this respect, we believe nEPS cell technology possess the potential of becoming an alternative strategy to generating insulin-producing cells in a relatively safe and efficient way. We were able to generate pancreatic β-cells that are functional, as proven by either in vitro or in vivo studies.
We have successfully developed a highly efficient differentiation protocols for human pluripotent stem cells that also yield functionally β-cells resembling those of adult human pancreatic β-cells. It showed the expression of insulin gene and protein in vitro
The transplanted nEPS-derived β-cells were able to survive within the tissue environment where they were engrafted. Moreover, preliminary data from a small cohort of DM mouse model (STZ-treated mice) also demonstrated a promising result in which correction of hyperglycemia was achieved following transplantation of nEPS-derived β-cells by IV injection and glucose level was decreased.
To further determine the functional status of insulin-producing cells, we analyzed glucose-dependent insulin release by ELISA. We found that beta-cell differentiated from nEPS released higher insulin levels in response to 27.7 mM glucose compared to 5.5 mM glucose (Figure). Beta-cell differentiated from nEPS showed higher insulin levels.
To estimate total cellular and secreted insulin levels, nEPS derived beta-cells were pre-incubated for 90 minutes at 37ºC in Krebs Ringer Bicarbonate Hepes (KRBH) buffer containing 118 mM sodium chloride, 4.7 mM potassium chloride, 1.1 mM potassium dihydrogen phosphate, 25 mM sodium hydrogen carbonate, 3.4 mM calcium chloride, 2.5 mM magnesium sulphate, 10 mM Hepes (all of sigma) and 2 mg/ml bovine serum albumin (Gibco) supplemented with 2.5 mM glucose. For induced insulin release, cells were further incubated in KRBH buffer supplemented with 27.7 mM glucose and alternatively with 5.5 mM glucose for 1h at 37°C and collected supernant. The control was incubated with 5.5 mM glucose. Supernatant samples containing secreted insulin were processed using the Human Ultrasensitive Insulin ELISA kit (Invitrogen). Cell numbers were counted automatically using Vi-Cell (Beckman Coulter).
Parkinson’s disease (PD) is a degenerative disorder of the central nervous system mainly affecting the motor system. Early in the course of the disease, the most obvious symptoms are movement-related; these include shaking, rigidity, slowness of movement and difficulty with walking and gait. Later, thinking and behavioral problems may arise, with dementia commonly occurring in the advanced stages of the disease, and depression being the most common psychiatric symptom.
nEPS homing effect in a Parkinson’s disease mouse model – Cell migration to substantia
nigra of mouse brain after nEPS injection through the tail vein
Stem cell research has the potential to significantly impact the development of disease-modifying treatments for Parkinson’s disease, and considerable progress has been made in creating dopamine-producing cells from stem cells. The development of new cell models of Parkinson’s disease is a particularly promising area of stem cell research, as the current lack of progressive, predictive models of Parkinson’s disease remains a major barrier to drug development. Cell models of Parkinson’s disease generated from stem cells could help researchers screen drugs more efficiently than in currently available animal models, and study the underlying biological mechanisms associated with Parkinson’s disease in cells taken from people living with the disease.
In 2013 PD was present in 53 million people and resulted in about 103,000 deaths globally. Parkinson’s disease is more common in older people, with most cases occurring after the age of 50; when it is seen in young adults, it is called young onset PD.