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Umbilical cord blood mesenchymal stem cell transplantation for the treatment of renal anemia:Hot spots and issues

Kai WANG ; Hongliang JIAO ; Jianbin LI ; Xianming WU

Chinese Journal of Tissue Engineering Research.2013;(32):5895-5900. doi:10.3969/j.issn.2095-4344.2013.32.023

BACKGROUND:Umbilical cord blood as a source of hematopoietic stem cel s has become a hot topic. Adjuvant therapy of umbilical cord blood transfusion has been used in China to correct renal anemia of uremia patients. OBJECTIVE:To evaluate the efficacy and safety of umbilical cord blood mesenchymal stem cel s in the treatment of renal anemia, and to compare with the effects of different sources of mesenchymal stem cel s with different transplantation methods on the treatment of kidney diseases. METHODS:A retrospective analysis was conducted on six renal anemia patients in the Department of Outpatient, Henan Red Cross Blood Center between January 2010 and December 2012. The experiment was approved by Medical Ethics Committee, six patients were informed consent for treatment programs, and parturients and their families have signed the informed consent. The newborns umbilical cord blood of 80-140 mL were col ected with closed sterile plastic blood bags, and then the separated umbilical cord blood mesenchymal stem cel s were transfused into the renal anemia patients through the superficial vein in the back of the hand with the number of≥1×108/copy, two copies per time, and re-transfused after 4 days, a total of three times. The blood hematocrit, hemoglobin, red blood cel s in urine, renal blood flow changes were observed before and after treatment. RESULTS AND CONCLUSION:The hemoglobin, blood hematocrit, red blood cel s in urine and renal blood flow were significantly increased before and after treatment (P<0.05). Transfusion of multiple copies of umbilical cord blood mesenchymal stem cel s through the superficial vein in the back of the hand is convenient and safe, which is considered as a new method for the treatment of renal anemia. But there are certain limitation in the data and the conclusion as the clinical research was designed as self-control, so further confirm is needed.

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Stem cell transplantation for ischemic heart disease:Clinical feasibility and safety

Yaguang HU

Chinese Journal of Tissue Engineering Research.2013;(32):5889-5894. doi:10.3969/j.issn.2095-4344.2013.32.022

BACKGROUND:The stem cel s transplanted into the damaged heart tissues can differentiate into cardiomyocytes, which bring new hope for the research of ischemic heart disease. OBJECTIVE:To explore the feasibility and safety of stem cel transplantation for the treatment of ischemic heart disease. METHODS:Various experimental methods to the feasibility and safety of stem cel transplantation for the treatment of ischemic heart disease were analyzed. REPAIR-AMI experiment was a randomized and double-blind and placebo-control ed multi-center study that used to analyze the therapeutic effect of intracoronary transplantation of bone marrow progenitor cel s immediately after acute myocardial infarction. MAGIC Cel-3-DES experiment was used to evaluate the safety of granulocyte colony-stimulating factor mobilized stem cel therapy and the effect of intracoronary injection of mobilized peripheral blood stem cel s on the treatment of acute myocardial infarction and old myocardial infarction. BOOST experiment was the randomized control ed study on autologous bone marrow cel transplantation through coronary vein after myocardial infarction. PROTECT-CAD experiment was the randomized control ed clinical trial about the direct injection of stem cel s into the myocardial for the treatment of ischemic heart disease. RESUTLS AND CONCLUSION:Stem cel transplantation may improve the left ventricular systolic function and left ventricular diastolic function as wel as the coronary flow reserve, and the related studies have been confirmed. Stem cel transplantation for the treatment of ischemic heart disease can increase the left ventricular ejection fraction. As the less clinical accident, stem cel therapy cannot increase the risk of restenosis based on the treatment of drug-eluting stents. It is safe and feasible of stem cel transplantation for the treatment of ischemic heart disease. Large sample, long scale and multi-center randomized control ed studies are needed to further evaluate the effect and risk of stem cel transplantation.

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Studies on p53 in induced pluripotent stem cells

Tongxiang LIN ; Yi LIN

Chinese Journal of Tissue Engineering Research.2013;(32):5883-5888. doi:10.3969/j.issn.2095-4344.2013.32.021

BACKGROUND:Induced pluripotent stem cel s can bypass the ethical issues of embryonic stem cel s, and become the hotspot of stem cel research. OBJECTIVE:To explore the research progress and problems of induced pluripotent stem cel s. METHODS:A retrospective analysis on the findings, research progress and problems of induced pluripotent stem cel s in recent years was performed. The Thomson Reuters Web of Science was searched for the articles related to the induced pluripotent stem cel s and p53 gene. RESULTS AND CONCLUSION:In recent years, domestic and foreign scholars have conducted a lot of researches on induced pluripotent stem cel s. For example, a Japanese group is establishing the stem cel bank to provide a basis for the treatment of retinal diseases. However, the safety issues of induced pluripotent stem cel s need to be solved before routine cel treatment application, in which the functional research of related p53 gene is one of the essential concerns. The other member of p53 gene, p73 gene, also participates in the generation and differentiation of induced pluripotent stem cel s, and the in-depth studies are needed. The finding of p53 gene function wil promote the in-depth development of regenerative medicine and translational medicine.

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Differentiation potential of umbilical cord blood-derived mesenchymal stem cells

Mingxing CHEN ; Guifang OUYANG

Chinese Journal of Tissue Engineering Research.2013;(32):5877-5882. doi:10.3969/j.issn.2095-4344.2013.32.020

BACKGROUND:Umbilical cord blood-derived mesenchymal stem cel s are multipotential stem cel s in the mesoderm in early development stage, and have been paid great attention due to its properties of multi-directional differentiation. OBJECTIVE:To summarize the potential of induced differentiation of umbilical cord blood-derived mesenchymal stem cel s. METHODS:We retrieved PubMed Database for articles concerning the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s published from January 1999 to December 2012. In titles and abstracts, the key words were“umblical cord blood, mesenchymal stem cel s, potential, differentiation”. Total y, 52 articles addressing the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s were reviewed. RESULTS AND CONCLUSION:Numerous studies have confirmed that human umbilical cord blood-derived mesenchymal stem cel s can successful y differentiate into multiple kinds of cel lines, but their understanding remains minor. If we can master the characteristics of the differentiation potential of umbilical cord blood-derived mesenchymal stem cel s, it would be used to repair bone and myocardium detects. Present studies remain in a starting stage. Isolation and purification, regulation of differentiation direction, in vitro amplification and immunogenicity require further investigations.

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Effect of bone marrow mesenchymal stem cells combined with Danhong injection on expression of GAP-43 and Bcl-2 after cerebral infarction

Jinyan LI ; Zhecheng ZHANG

Chinese Journal of Tissue Engineering Research.2013;(32):5871-5876. doi:10.3969/j.issn.2095-4344.2013.32.019

BACKGROUND:Danhong injection, scavenging free radicals and inhibiting lipid peroxidation, can improve microenvironment injury after cerebral infarction. OBJECTIVE:To explore the influence of bone marrow mesenchymal stem cel s combined with Danhong injection on expression of GAP-43 and Bcl-2 after cerebral infarction in rats. METHODS:Sixty Wistar rats were selected to prepare models of cerebral infarction by middle cerebral artery occlusion and then randomly divided into control group, bone marrow mesenchymal stem cel group, and combination group. Control group received tail vein injection of PBS. Bone marrow mesenchymal stem cel group received tail vein injection of 2.5×109/L bone marrow mesenchymal stem cel suspension. Combination group received injection of 2.5× 109/L bone marrow mesenchymal stem cel suspension+2 mL/kg Danhong injection, for 5 consecutive days, once a day. RESULTS AND CONCLUSION:There were no significant differences in the neurological dysfunction scores among the three groups at 24 hour and 3 days after implantation (P>0.05). The neurological dysfunction scores in the ombination group were significantly lower than those in the bone marrow mesenchymal stem cel group and control group at 1 and 2 weeks after transplantation (P<0.05). In the combination group, GAP-43 and Bcl-2 expression was significantly higher than the bone marrow mesenchymal stem cel group and control group (P<0.05). Bone marrow mesenchymal stem cel transplantation combined with Danhong injection can significantly promote the local expression of GAP-43 and Bcl-2 after cerebral infarction, and has obvious inhibitory effects on cel apoptosis in rats with cerebral infarction.

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Tougu Xiaotong Granule containing serum induces cartilage differentiation of bone marrow stromal stem cells

Boling LIU ; Xihai LI ; Lichun XIAO ; Guiqing LIANG ; Guangwen WU ; Zhaohui LI ; Qiyong CHEN

Chinese Journal of Tissue Engineering Research.2013;(32):5863-5870. doi:10.3969/j.issn.2095-4344.2013.32.018

BACKGROUND:Bone marrow stromal stem cel s have multilineage differentiation capacity and can differentiate into transparent chondrocytes under certain conditions, which can provide new thoughts for treatment of osteoarthritis. OBJECTIVE:To investigate the effects of Tougu Xiaotong Granule containing serum on the cartilage differentiation of bone marrow stromal stem cel s. METHODS:Bone marrow stromal stem cel s from Sprague-Dawley rat limbs were cultured in vitro, and those cel s at passage 3 were used in the study. Cel s were divided into six groups:saline serum group, Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group. The Sox9, col agen Ⅱ, and col agen X mRNA and protein expression levels were tested by real-time polymerase chain reaction and western blot analysis. RESULTS AND CONCLUSION:After cel s were intervened with drug-containing serum for 14 days, the Sox9, col agen Ⅱ, and col agen X mRNA and protein expression in Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group were significantly higher than that in saline serum group (P<0.05, P<0.01). In addition, the chondroinductive group, Tougu Xiaotong Granule water extract serum and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group showed significantly higher expression levels than Tougu Xiaotong Granule water extract serum group, Tougu Xiaotong Granule alcohol extract group (P<0.01). Sox9 expression in Tougu Xiaotong Granule water extract serum and chondroinductive group were significantly higher than that in the chondroinductive group. Experimental findings indicate that, Tougu Xiaotong Granule containing serum can accelerate bone marrow stromal stem cel s differentiate into cartilage cel s by up-regulation of Sox9 expression.

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Periodontal ligament stem cells expansion in vitro under different cryopreservation systems

Xuan WANG ; Yishan LIU ; Yan MA ; Xiaojuan BI ; Shutao ZHENG ; Jia LIU ; Boqi LI ; Dalei SUN ; Jin ZHAO

Chinese Journal of Tissue Engineering Research.2013;(32):5855-5862. doi:10.3969/j.issn.2095-4344.2013.32.017

cryopreservation periodontal tissue. METHODS:Periodontal tissue was scraped from healthy human teeth and divided them into three equal parts:fresh group, harvesting periodontal ligament stem cel s from fresh tissue;5%dimethyl sulfoxide group, 5%dimethyl sulfoxide added into the cryopreservation system;10%dimethyl sulfoxide group, 10%dimethyl sulfoxide added into the cryopreservation system. One month later, periodontal ligament stem cel s were extracted from the cryopreserved periodontal ligament from the latter two groups. RESULTS AND CONCLUSION:In the time that cel s swam out of tissue mass and cel harvest amount, the 5%dimethyl sulfoxide group was inferior to the fresh group but better than the 10%dimethyl sulfoxide group (P<0.05). No differences were found among the three groups in the fol owing aspects (P>0.05):colony formation rate of passage 1 periodontal ligament stem cel s, cel survival rate, proliferation ability of passage 3 periodontal ligament stem cel s, cel growth curve and surface marker expression of periodontal ligament stem cel s. The results suggest that the 5%dimethyl sulfoxide added cryopreserved system for periodontal ligament tissue cannot only shorten the time of periodontal ligament stem cel s amplification in vitro, ensure cel harvest and maintain basic cel ular biological characteristics, but also reduce the total amount of the dimethyl sulfoxide and the direct damage to the cel s caused by repeatedly frozen cel s, thereby providing a more secure guarantee for the future implementation of clinical transplantation therapy. So, the 5%dimethyl sulfoxide added cryopreserved system may be the new selection for donor tissue storage.

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Evaluation indexes for the viability of umbilical cord-derived mesenchymal stem cells before transplantation

Xin LEI ; Yan CHEN ; Jianlin ZHANG ; Lei CUI ; Yuhu NIU ; Bo NIU

Chinese Journal of Tissue Engineering Research.2013;(32):5847-5854. doi:10.3969/j.issn.2095-4344.2013.32.016

BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.

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Low-frequency electromagnetic fields enhance the recovery of spinal cord injured rats undergoing bone mesenchymal stem cell transplantation

Yu FENG ; Wenfang BAI ; Weicheng XU ; Xinping LI ; Liming BAI ; Ling LIANG ; Xin WANG ; Mingsheng ZHANG

Chinese Journal of Tissue Engineering Research.2013;(32):5819-5826. doi:10.3969/j.issn.2095-4344.2013.32.012

BACKGROUND:Bone marrow mesenchymal stem cel transplantation is considered as a promising therapy for spinal cord injury. How to more effectively promote the survival of bone marrow mesenchymal stem cel s in the area of spinal cord injury and to accelerate the recovery of motor function after spinal cord injury is a current study focus. Previous studies have found that low-frequency electromagnetic fields can promote bone marrow mesenchymal stem cel proliferation and differentiation, but whether the low-frequency electromagnetic fields can be applied to bone marrow mesenchymal stem cel transplantation for treatment of spinal cord injury requires further studies. OBJECTIVE:To discuss the effects of low-frequency electromagnetic fields on motor function of spinal cord injury rats after transplantation of bone mesenchymal stem cel s. METHODS:Sixty-four rat models of incomplete spinal cord injury at T 10 were established by compression method and then randomized into control group, transplantation group (bone mesenchymal stem cel transplantation), electromagnetic field group and combination group (electromagnetic field+bone mesenchymal stem cel transplantation). After successful modeling, bone mesenchymal stem cel s labeled with 5-bromo-2'-deoxyuridine were injected into the original injured site in the transplantation group and combination group, which were isolated and purified with the fast adherence method;while alpha-minimum essential medium was injected into the electromagnetic field group and control group for instead. At 24 hours post-operation, the electromagnetic field group and combination group were explored to low-frequency electromagnetic fields (frequency 50 Hz, magnetic indaction intensity 5 mT) for 60 minutes per day. RESULTS AND CONCLUSION:After cel transplantation for 21 days, the Basso, Beattie, and Bresnahan scores in the combination group was higher than the other groups (P<0.05). 5-Bromo-2'-deoxyuridine positive cel s grew wel , and integrated into the normal spine;syringomyelia was reduced, and the number of spinal neural cel s was increased in the combination group. In addition, glial fibril ary acidic protein expression was decreased in the combination group, while matrix metal oproteinase 2 expression was increased. It indicates that low-frequency electromagnetic fields could promote recovery of motor function in the spinal cord injury rats transplanted with bone mesenchymal stem cel s, which could be associated that low-frequency electromagnetic fields facilitate the survival of transplanted bone mesenchymal stem cel s, up-regulate the expression of matrix metal oproteinase 2, and reduce glial scar formation in the spinal cord injured site.

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Mouse embryonic hepatic stem cells differentiate into cardiomyocyte-like cells

Bigang WU ; Jing CHANG ; Xiaogang ZHANG

Chinese Journal of Tissue Engineering Research.2013;(32):5813-5818. doi:10.3969/j.issn.2095-4344.2013.32.011

BACKGROUND:In recent years, embryonic hepatic stem cel s have attracted more attention, but there are few reports on the potential of embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s as wel as the related differentiation conditions. OBJECTIVE:To investigate the moderate condition to induce mice embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s in vitro with chemical reagents. METHODS:Dimethylsulfoxide in combination with 5-azacytidine with different concentrations and time were used to induce the embryonic hepatic stem cel s of 13.5 days mice and to observe the differentiation effect. RESULTS AND CONCLUSION:Under in vitro conditions, 0.8%dimethylsulfoxide+5μmol/L 5-azacytidine could induce the mouse embryonic hepatic stem cel s to express the specific markers of myocardial cel s, while increasing the concentration of the inducer and extending the induction time could not improve the induction efficacy.

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