ABSTRACT
Human adipose stem cells (hASCs) were introduced as appropriate candidate due to advantages like ease of isolation, in vitro expansion and lack of immune response. Deprenyl (Dep) was used to induce bone marrow stem cells into neuron-like cells. We investigated the Dep effect on neurotrophin genes expression in hASCs and their differentiation into neuron-like cells. The cells were isolated from small pieces of abdominal adipose tissue and subjected to flow cytometry to confirm purification. The osteogenic and adipogenic differentiation were identified. The proliferation rate and neurotrophin genes expression of treated cells were evaluated by MTT, TH immunostaining and RT-PCR. hASCs had positive response to CD44, CD73, CD90, CD105 markers and negative response to CD34 and CD45 markers and differentiated into adipocytes and osteocytes. Exposure to 10–7 M of Dep for 24 hours caused a significant increase of viable cells and BDNF, NTF-3 genes expression as compared to cultured cells in serum free medium and had no effect on the expression of NGF and GDNF genes. Based on our results, Dep is able to induce BDNF, NTF-3 and NTF-4 genes expression and neroun-like morphology in hASCs.
ABSTRACT
Human adipose stem cells (hASCs) were introduced as appropriate candidate due to advantages like ease of isolation, in vitro expansion and lack of immune response. Deprenyl (Dep) was used to induce bone marrow stem cells into neuron-like cells. We investigated the Dep effect on neurotrophin genes expression in hASCs and their differentiation into neuron-like cells. The cells were isolated from small pieces of abdominal adipose tissue and subjected to flow cytometry to confirm purification. The osteogenic and adipogenic differentiation were identified. The proliferation rate and neurotrophin genes expression of treated cells were evaluated by MTT, TH immunostaining and RT-PCR. hASCs had positive response to CD44, CD73, CD90, CD105 markers and negative response to CD34 and CD45 markers and differentiated into adipocytes and osteocytes. Exposure to 10–7 M of Dep for 24 hours caused a significant increase of viable cells and BDNF, NTF-3 genes expression as compared to cultured cells in serum free medium and had no effect on the expression of NGF and GDNF genes. Based on our results, Dep is able to induce BDNF, NTF-3 and NTF-4 genes expression and neroun-like morphology in hASCs.
ABSTRACT
There is longstanding experimental and clinical evidence that supports the idea that replacement of dopaminergic [DAergic] neurons can ameliorate functional disabilities of Parkinson's disease [PD]. The purpose of the present study is to examine the efficacy of transplantation of rat bone marrow stromal cell [BMSCs]-derived tyrosine hydroxylase-positive [TH[+]] cells induced by deprenyl into 6-hydroxydopamine [6-OHDA]-lesioned rat models, using behavioral tests and immunohistochemical evaluations. In this experimental study, undifferentiated BrdU-labeled BM-SCs were incubated in serum-free medium that contained 10[-8] M deprenyl for 24 hours. Afterwards, BMSCs were cultured for 48 hours in alpha-minimal essential medium [alpha-MEM] supplemented with 10% FBS, then differentiated into Th[+] neurons. We randomly divided 24 hemiparkinsonian rats as follows: group 1 [control] received only medium, while groups 2 and 3 were injected with 2x10[5] BMSCs and deprenyl-treated cells in 4 microl medium. Injections were made into the injured strata of the rats. Rotational behavior in response to apomorphine was tested before transplantation and at 2, 4, and 6 weeks post-graft. Animals were then sacrificed, and the brains were extracted for immunohistochemical and electron microscopic studies. Apomorphine-induced rotation analysis indicated that animals with grafted cells in groups 2 and 3 exhibited significantly less rotational behavior than those in the control group at 2, 4, and 6 weeks after transplantation. Immunohistochemical analysis demonstrated that BrdU-labeled cells expressed specific neuronal markers, such as NF 200 and TH, at the implantation site. The presence of TH[+] cells in conjunction with the reduction in rotation might show the capacity of grafted cells to release dopamine. Ultrastructural analysis revealed the presence of immature neurons and astrocyte-like cells at the graft site. Th[+] neurons induced by deprenyl can be considered as a cell source for PD autograft therapy?
Subject(s)
Male , Animals, Laboratory , Selegiline , Tyrosine 3-Monooxygenase , Oxidopamine , Rats , Models, Animal , Parkinson Disease , Immunohistochemistry , BehaviorABSTRACT
It has been reported that rat bone marrow stromal cells [BMSCs] can be spontaneously differentiated into neural-like cells without any supplemental growth factors and/or chemical treatment after long-term culture. This study aims to determine whether, growth factors secreted by MSCs could induce self-differentiation into neural-like cells in a long-term culture. This study consisted of two groups: i. rat BMSCs [passage 5] were cultured in alfa- minimal essential medium [alpha-MEM] and 10% fetal bovine serum [FBS] without the addition of inducer and exchanging medium for three weeks, as the experimental group and ii.rat BMSCs [passage 5] as the control group. Each group was analysed by reverse transcriptase polymerase chain reaction [RT-PCR] to evaluate the expressions of neurotrophic factors and neural marker genes. Statistical analyses were carried out using one-way analysis of variance [ANOVA] and Tukey's multiple comparison with SPSS software [version 16]. P< 0.05 was considered statistically significant. The experimental group [fifth passage of BMSCs] obtained from adult rats spontaneously differentiated into neural precursor cells after long-term culture. Cultured cells expressed tyrosine hydroxylase [TH], Nurr1 and nestin genes. Furthermore, some growing cells in suspension became neurosphere-like. Self-differentiated rat MSCs [SDrMSCs] expressed significantly higher levels of NGF [0.96 +/- 0.16], nestin [0.63 +/- 0.08], and Nurr1 [0.80 +/- 0.10] genes [p<0.05]. In this study, we reported that rMSCs in long-term culture underwent spontaneous transformation to neural precursors without the supplement of growth factors and specific chemicals. Cells expressed neural markers such as: TH, Nurr1, and nestin genes