Management of intrauterine adhesions using human amniotic mesenchymal stromal cells to promote endometrial regeneration and repair through Notch signalling.

Intrauterine adhesions (IUAs) severely hamper women's reproductive functions. Human amniotic mesenchymal stromal cell (hAMSC) transplantation is effective in treating IUAs. Here, we examined the function of Notch signalling in IUA treatment with hAMSC transplantation. Forty-five Sprague-Dawley female rats were randomly divided into the sham operation, IUA, IUA + E2, IUA + hAMSCs and IUA + hAMSCs + E2 groups. After IUA induction in the rats, hAMSCs promoted endometrial regeneration and repair via differentiation into endometrial epithelial cells. In all groups, the expression of key proteins in Notch signalling was detected in the uterus by immunohistochemistry. The results indicated Notch signalling activation in the hAMSCs and hAMSCs + E2 groups. We could also induce hAMSC differentiation to generate endometrial epithelial cells in vitro. Furthermore, the inhibition of Notch signalling using the AdR-dnNotch1 vector suppressed hAMSC differentiation (assessed by epithelial and mesenchymal marker levels), whereas its activation using the AdR-Jagged1 vector increased differentiation. The above findings indicate Notch signalling mediates the differentiation of hAMSCs into endometrial epithelial cells, thus promoting endometrial regeneration and repair; Notch signalling could have an important function in IUA treatment.

The study for induction and differentiation of rat's adipose-derived stem cells by valproic acid in vitro / 中华显微外科杂志

Objective To study the inducting differentiation effects of the vaproic acid (VPA) on rats adipose derived stem cells (ADSCs) in vitro.Methods From November,2016 to October,2017,the ADSCs were isolated from 2 healthy 3-weeks-old Sprague-Dawley (SD) rats and cultured to passage 3,which were treated with 10 ng/ml bFGF for 24 hours before induction.Then the induction media contained the VPA with different concentrations:group B (0.5 mmol/L,200 μl) VPA;group C (1.0 mmol/L,200 μl) VPA;group D (10 mmol/L,200 μl) VPA,and D-Hank was used in group A as blank control group.The morphological changes of the cells were observed every day.At 7 days of induction,the gene expressions of neuron-specific enolase (NSE),nestin (NES),and S-100 were detected by real-time fluorescent quantitative PCR.The S-100 protein expression was tested by immunofluorescence staining.Significance of difference was determined using independent t test.Probabilities lower than 5％ (P ＜ 0.05) were considered statistically significant.Results At 4 days after induction,some ADSCs of groups B,C,and D showed the morphology of Schwann-like cells or neuron-like cells,the change of group C was more obvious;and the AD-SCs of group A had no obvious change,which still present spindle.The S-100 immunofluorescence staining showed higher ratio of positive expression in groups B,C,and D (more obvious in group C) and lower ratio of positive expression in groups A and D (more obvious in group A).The gene expression of S-100 showed dose-dependent increases in groups C,which was significantly higher than that of groups A,B,and D (P＜0.05),but no significant differ ence was found between groups B and D (P＞0.05).The gene expression of NSE showed the same tendency as S-100,which reached the peak in group C;the gene expression of NSE in group C was significantly higher than that of groups A,B,and D (P＜0.05),and groups B and D showed significant difference (P＜0.05).However,the expression of Nestin showed no significant difference among these groups (P＞0.05).Conclusion ADSCs can be induced to differentiate into Schwann-like cells or neuron-like cells under the treatment of VPA,and 1.0 mmol/L is the optimal concentration.

Different ingredients of cell culture medium influence multi-differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.23.017

Detection of Ca2+i in neuron-like cells from bone mesenchymal stem cell differentiation induced by salvia miltiorrhiza / 中华神经医学杂志

Objective To measure the concentration of intracellular free Ca2+ ([Ca2+]i) in neuron-like cells resulted from rat bone mesenchymal stem cell (BMSCs) differentiation induced by salvia miltiorrhiza injection and provide some theoretical basis for the BMSCs transplantation. Methods The rat BMSCs were separated from rat bone marrow and cultured in vitro. After induced by basic fibroblast growth factor and 10mL/L salvia miltiorrhiza injection, the cells were identified with immunofluorescence staining against NeuN. The same procedure was performed on primarily cultured hippocampal neurons. Then, the [Ca2+]i of the differentiated neuron-like cells was determined and compared with primarily cultured hippocampal neurons. Results The BMSCs after induced by basic fibroblast growth factor and salvia miltiorrhiza injection expressed neuronal phenotypes similar to the cell appearance of neurons with NeuN. The average fluorescence intensity of the neuron-like cells derived from BMSCs was 984.75±79.51, while the average fluorescence intensity of the primarily cultured hippocampal neurons was 769.42±60.93. No significant difference was found between them (P>0.05). Conclusion The neuron-like cells from rat BMSCs differentiation induced by salvia miltiorrhiza injection possess certain neuronal properties.

Differentiation of rat bone marrow stromal cells into Schwann cells-like in vitro / 中华显微外科杂志

Objective To study the protocol that rat bone marrow stromal cells (BMSCs) can be induced to differentiate into Schwann cell-like in vitro. Methods BMSCs were treated with beta-mercaptoethanol followed by retinoic acid and cultured in the presence of forskolin,basic-FGF,PDGF and heregulin. The positive percentages of GFAP,S-100 protein expression were measured by immunocytochemistry SABC staining. Results After the induction,BMSCs displayed morphologies of Schwann cell,such as ellipsoidal cell bodies and fences-like array,with two or three sligh cell processes. The positive percentages of GFAP protein expression was from 42.5% to 68.7%,S-100 was from 39.6% to 60.2%. Conclusions Rat BMSCs could be induced to Schwann cell-like by beta-mercaptoethanol,retinoic acid,forskolin,basic-FGF,PDGF and heregulin

The specialization of mesenchymal stem cells to cardiac muscle cells with trichostatin A intervention in the simulated microenvironment / 重庆医科大学学报

Objective:This study was to investigate the interrelations between the acetylase activity and specialization rate of the differentiation of MSCs to the cardiac cells when histon deacetylase enzyme inhibitor trichostatin A (TSA)interferes in the cardiac cells. Method: MSCs of ratswere separated,The already labeled MSCs and cardiac muscle cells were put respectively into the MSCs interfered by different concentrations of TSA cultivated in mix tune with cardiac muscles. Adding TSA into the conventional culture of MSCs for the negative group.One week later,with the help of immunofluorescence,detecting the expression of the function proteinum,ie. myosin heavy chain (MHC)and Connexin43 (Cx43),of the cardiac muscle constitution in MSCs was detected. Results: ① There appeared sample differentiation of the cardiac muscle cells after the coculture of the MSCs and the pulsating cardiac muscle cells for one week,and the expression of the function proteinum of the cardiac muscle constitution;② In the positive group,in the mix culture of the cardiac muscle cells and the MSCs in which the TSA interfered for 48 h,there appeard sample differentiation of the cardiac muscle cells;③ In the negative group,part of the MSCs could express the function proteinum of the cardiac muscle constitution. During the 7 days of coculture, with the induction of TSA,MSCs was double-labeled by Brdu-DAPI. The differentiation rate of the fluorescent manifestation of the MHC and Cx43 in the MSCs was notably different expression that in the positive group,Conclusion: MSCs in which HDAC,the enzyme inhibitor,interferes are in the microenvironment of the cardiac muscle cells. HDAC,the enzyme inhibitor,functions as a promotion factor in the specialization of MSCs,suggesting the interrelation between the acetylase activity and the cell specialization rates.