8 research outputs found
In vitro evaluation of different protocols for the induction of mesenchymal stem cells to insulin-producing cells
Stem cells therapy is a new promising approach for
diabetes mellitus (DM) treatment, but the insulin secretion
rate in differentiated cells is low when compared with pancreas
beta cells embedded in Langerhans islets. In this study,
we evaluated different protocols of insulin secretion to
achieve the most appropriate protocol for in vitro insulin
secretion. We differentiated human umbilical cord matrixderived
mesenchymal cells (hUCMs) into insulinproducing
cell (IPC) by the aim of three previously reported
protocols and a modified protocol. The insulin content was
analyzed through gene expression and immunocytochemistry
(IHC). Dithizone (DTZ) staining was done for identification
of islet-like structures. Insulin and C peptide secretion
was measured by chemiluminesence immunoassay (CLIA)
and enzyme immunoassay (EIA) as well. Reverse
transcription-PCR (RT-PCR) showed efficient expression of
insulin genes in all the study groups. IHC analysis showed
higher expression of insulin and proinsulin proteins in the
modified protocol. DTZ staining exhibited variable islet-like
clusters in the different protocols except control. This finding
was confirmed by the higher response to glucose challenge
test in this group. A modified protocol using an intermediate
step that makes the cells vulnerable to nestin production
in combination with inducing agent results in the
higher differentiation of stem cells into insulin-producing
cells and more insulin secretion in vitro
Transplantation of differentiated umbilical cord mesenchymal cellsunder kidney capsule for control of type I diabetes in rat
Nowadays, stem cells have been introduced as an appropriate source of regenerative medicine fortreatment of type I diabetes. Human umbilical cord matrix-derived mesenchymal cells (hUCMC) havesuccessfully been differentiated into insulin producing cells. The isolated hUCM cells were characterizedby the expression of stem cell surface markers and by differentiation into adipocytes and osteocytes.The hUCMCs were cultured with different concentrations of neural conditional medium (NCM) and wereinduced to differentiate into insulin producing cells (IPCs). As 60% NCM concentration resulted in highernestin and PDX1 expression, the cells were first exposed to 60% NCM and were then induced for IPCsdifferentiation. PDX1 and insulin gene expression was evaluated in the treated cells. Also, the secretioncapacity of the IPCs was assessed by glucose challenge test. IPCs were transferred under the rat kid-ney capsule. Blood glucose level, weight gain and immunohistochemistry assessments were done in thetreated animals. hUCMC expressed mesenchymal cell surface markers and successfully differentiatedinto adipocytes and osteocytes. Higher NCM concentration resulted in higher PDX1 and nestin expres-sion. The IPCs expressed insulin and PDX1. IPCs were detectable under the kidney capsule 2 monthsafter injection. IPCs transplantation resulted in a sharp decline of blood sugar level and less weight loss.Differentiated hUCM cells could alleviate the insulin deprivation in the rat model of type I diabetes. Inaddition, higher NCM concentration leads to more differentiation into IPCs and more nestin and PDX1expression. Kidney capsule can serve as a suitable nominee for IPCs transplantation
Effects of transplanted mesenchymal stem cells isolated from Wharton’s jelly of caprine umbilical cord on cutaneous wound healing; histopathological evaluation
The aim of this study was to investigate the effects of transplanted Wharton’s
jelly mesenchymal stem cells (WJMSCs) of caprine umbilical cord on cutaneous wound
healing process in goat. After collection of caprine pregnant uterus of mixed breed goats
from abattoir, the Wharton’s jelly (WJ) of umbilical cord was harvested. The tissues were
minced in ventilated flasks and explant culture method was used for separating
mesenchymal stem cells (MSCs). The isolated cells were immunostained for Actin protein,
histochemically assayed for the presence of alkaline phosphatase activity, and analyzed for
detection of matrix receptors (CD44) and hematopoetic lineage markers (CD34), using flow
cytometery. After The isolated cells, 3×106 MSCs were stained with BrdU and prepared for
transplantation to each wound. Four 3-cm linear full thickness skin incisions were made on
both sides of thoracic vertebrate of four Raeini goats (two wounds on each side). The left
wounds were implanted with MSCs in 0.6 ml of Phosphate buffer saline (PBS), and the
right wounds considered as control group that received 0.6 ml of PBS. The samples were
taken from the wounds 7 and 12 days after the wounding, and healing process was
compared histologically between the two groups. Anti-BrdU staining showed that the
transplanted cells were still alive in the wound bed during the study. The histopathological
study revealed that re-epithelialization was complete at days 7 in treated wounds with
WJMSCs, whereas in control wound the wounds still showed incomplete epithelialization 12 days after wounding. Also, microscopic evaluation showed less inflammation, thinner
granulation tissue formation with minimum scar in the treated wounds in comparison with
control wounds. In conclusion, this study demonstrates the beneficial effect of caprine
WJMSCs in cutaneous wound healing in goat
Neural differentiation of human umbilical cord matrix-derived mesenchymal cells under special culture conditions
Many neural disorders are characterized by the loss of one or several types of neural cells. Human umbilical cord-derived mesenchymal cells (hUCMs) are capable of differentiating into neuron, astroglia-like and oligodendrocyte cell types. However, a reliable means of inducing the selective differentiation of hUCMs into neural cells in vitro has not yet been established. For induction of neural differentiation, hUCMs were seeded onto sterile glass slides and six various cocktails using a base medium (DMEM/LG) supplemented with 10 % FBS, retinoic acid (RA), dimethyl sulfoxide (DMSO), epidermal growth factor (EGF) and fibroblast growth factor (FGF) were used to compare their effect on neuronal, astrocyte and oligodandrocyte differentiation. The hUCMs were positive for mesenchymal markers, while they were negative for hematopoietic markers. Differentiation to adipogenic and osteogenic lineage was detected in these cells. Our data revealed that the cocktail consisting of DMEM/LG, FBS, RA, FGF, and EGF (DF/R/Fg/E group) induced hUCM cells to express the highest percentage of nestin, ß-tubulin III, neurofilament, and CNPase. The DF/Ds/Fg/E group led to the highest percentage of GFAP expression. While the expression levels of NF, GFAP, and CNPase were the lowest in the DF group. The least percentage of nestin and ß-tubulin III expression was observed in the DF/Ds group. We may conclude that FGF and EGF are important inducers for differentiation of hUCMs into neuron, astrocyte and oligodendrocyte. RA can induce hUCMs to differentiate into neuron and oligodendrocyte while for astrocyte differentiation DMSO had a pivotal role
Neural differentiation of human umbilical cord matrixderived mesenchymal cells under special culture conditions
Many neural disorders are characterized
by the loss of one or several types of neural cells.
Human umbilical cord-derived mesenchymal cells
(hUCMs) are capable of differentiating into neuron,
astroglia-like and oligodendrocyte cell types. However,
a reliable means of inducing the selective
differentiation of hUCMs into neural cells in vitro
has not yet been established. For induction of neural
differentiation, hUCMs were seeded onto sterile glass
slides and six various cocktails using a base medium
(DMEM/LG) supplemented with 10 % FBS, retinoic
acid (RA), dimethyl sulfoxide (DMSO), epidermal
growth factor (EGF) and fibroblast growth factor
(FGF) were used to compare their effect on neuronal,
astrocyte and oligodandrocyte differentiation. The
hUCMs were positive for mesenchymal markers,
while they were negative for hematopoietic markers.
Differentiation to adipogenic and osteogenic lineage
was detected in these cells. Our data revealed that the
cocktail consisting ofDMEM/LG, FBS, RA, FGF, and
EGF (DF/R/Fg/E group) induced hUCM cells to
express the highest percentage of nestin, ß-tubulin
III, neurofilament, and CNPase. The DF/Ds/Fg/E
group led to the highest percentage of GFAP expression.
While the expression levels of NF, GFAP, and
CNPase were the lowest in the DF group. The least
percentage of nestin and ß-tubulin III expression was
observed in the DF/Ds group. We may conclude that
FGF and EGF are important inducers for differentiation
of hUCMs into neuron, astrocyte and oligodendrocyte.
RA can induce hUCMs to differentiate into
neuron and oligodendrocyte while for astrocyte
differentiation DMSO had a pivotal role
The effect of aqueous extract of Rosa damascena on formaldehyde-induced toxicity in mice testes
Context: Rosa damascena L. (Rosaceae) (RD) essential oil and extracts are commonly used as a flavour in herbal medicine which increase libido. Previous studies have shown inhalation of RD flower’s oil increases libido and causes protective effects in formaldehyde (FA)-induced testicular damage. Objective: The protective effects of aqueous extract of RD on the male reproductive system of mice were examined following FA-induced damage. Materials and methods: Forty-eight adult NMRI male mice were randomly assigned to six groups (n = 8): control (normal saline, 10 mg/kg); RD40 (40 mg/kg, p.o.); FA treated (10 mg/kg of 10%, i.p.) and FA + RD treated at 10, 20 and 40 mg/kg (FA + RD10), (FA + RD20) and (FA + RD40), respectively, for 40 days. At the end of treatment regimes, serum testosterone (T) level and the reproductive activity, viz. body/organ weights, testicular structure and sperm characteristics were studied. Results: Formaldehyde administration significantly decreased serum T level (p < 0.001), testicular weight/volume, tubular diameter and sperm characteristics compared to the control group (p < 0.05). RD (40 mg/kg) administration in FA-treated mice significantly improved serum T level, testicular weight/histological structure, tubular diameter, Leydig cell number and epididymal sperm characteristics in comparison to its lower doses and the control group (p < 0.05). Discussion and conclusions: We may conclude that RD flower extract can withstand effects of FA in the male reproductive system of mice possibly due to its antioxidative properties