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1.
A novel, minimally invasive, transabdominal embryo collection method (transabdominal method) was developed as an alternative to a standard abdominal incision for embryo collection in the common marmoset. The abdominal incision method was used for 304 flushes using 36 female animals, whereas the transabdominal method was used for 488 flushes using 48 females; successful embryo collection rates were 48.0% and 48.4% (P > 0.05), respectively. These techniques were successfully duplicated at another institute (German Primate Center, DPZ). At that institution, successful embryo collection rates were 88.9% and 77.8% for the abdominal incision and transabdominal methods, respectively (P > 0.05), whereas the average numbers of preimplantation embryos obtained per flush were (mean ± SD) 1.91 ± 0.35 and 1.71 ± 0.14 (P > 0.05). The transabdominal method reduced animal stress, did not require incisional wound healing, and enabled successive embryo recoveries to be done much sooner. More embryos in early developmental stages (zygotes/morulae) were recovered using the transabdominal method (76.1%) than the abdominal incision method (52.6%, P < 0.01). In contrast, recovery of arrested or abnormal embryos was not significantly different between these two methods (9.8% and 8.3%). To verify developmental ability of embryos recovered by the transabdominal method, transfer of 28 normal embryos to 14 surrogate mothers yielded a nidation rate of 57%. Five females sustained term pregnancies and eight neonates were born. This novel transabdominal method will facilitate progress in marmoset developmental biology and embryology. 相似文献
2.
Nobuhiro Shimozawa Ryoichi Ono Manami Shimada Hiroaki Shibata Ichiro Takahashi Hiroyasu Inada Tatsuyuki Takada Tetsuya Nosaka Yasuhiro Yasutomi 《Differentiation; research in biological diversity》2013,85(4-5):131-139
Induced pluripotent stem (iPS) cells established by introduction of the transgenes POU5F1 (also known as Oct3/4), SOX2, KLF4 and c-MYC have competence similar to embryonic stem (ES) cells. iPS cells generated from cynomolgus monkey somatic cells by using genes taken from the same species would be a particularly important resource, since various biomedical investigations, including studies on the safety and efficacy of drugs, medical technology development, and research resource development, have been performed using cynomolgus monkeys. In addition, the use of xenogeneic genes would cause complicating matters such as immune responses when they are expressed. In this study, therefore, we established iPS cells by infecting cells from the fetal liver and newborn skin with amphotropic retroviral vectors containing cDNAs for the cynomolgus monkey genes of POU5F1, SOX2, KLF4 and c-MYC. Flat colonies consisting of cells with large nuclei, similar to those in other primate ES cell lines, appeared and were stably maintained. These cell lines had normal chromosome numbers, expressed pluripotency markers and formed teratomas. We thus generated cynomolgus monkey iPS cell lines without the introduction of ecotropic retroviral receptors or other additional transgenes by using the four allogeneic transgenes. This may enable detailed analysis of the mechanisms underlying the reprogramming. In conclusion, we showed that iPS cells could be derived from cynomolgus monkey somatic cells. To the best of our knowledge, this is the first report on iPS cell lines established from cynomolgus monkey somatic cells by using genes from the same species. 相似文献
3.
G. E. Webley D. H. Abbott L. M. George J. P. Hearn H. Mehl 《American journal of primatology》1989,17(1):73-79
This study describes the concentrations of melatonin in plasma samples taken from marmoset monkeys (Callithrix jacchus) every 4 h over three 24-h periods. A circadian pattern of secretion was apparent, with higher levels recorded at night (20.00–08.00 h) than during the day (08.00–20.00 h) and a peak concentration at 20.00 h. There was a significant difference in the mean day and night concentrations (32.5 ± 4.5 pg/ml versus 49.0 ± 6.9 pg/ml, respectively) with individual concentrations ranging between<10–60 pg/ml in the day and 15–200 pg/ml at night. Circadian plasma melatonin concentrations were similar over the three 24-h periods, in male (n = 3) and female (n = 3) monkeys, and in dominant (cyclic, n = 5) and subordinate (acyclic, n = 4) females. The results show a less pronounced circadian profile in the marmoset than is seen in the human but a similar profile to that in the seasonally breeding rhesus monkey. 相似文献
4.
The regeneration potential of mesenchymal stem cells (MSCs) diminishes with advanced age and this diminished potential is associated with changes in cellular functions. This study compared MSCs isolated from the bone marrow of rhesus monkeys (rBMSCs) in three age groups: young (< 5 years), middle (8-10 years), and old (> 12 years). The effects of aging on stem cell properties and indicators of stem cell fitness such as proliferation, differentiation, circadian rhythms, stress response proteins, miRNA expression, and global histone modifications in rBMSCs were analyzed. rBMSCs demonstrated decreased capacities for proliferation and differentiation as a function of age. The production of heat shock protein 70 (HSP70) and heat shock factor 1 (HSF1) were also reduced with increasing age. The level of a core circadian protein, Rev-erb α, was significantly increased in rBMSCs from old animals. Furthermore, analysis of miRNA expression profiles revealed an up-regulation of mir-766 and mir-558 and a down-regulation of mir-let-7f, mir-125b, mir-222, mir-199-3p, mir-23a, and mir-221 in old rBMSCs compare to young rBMSCs. However, there were no significant age-related changes in the global histone modification profiles of the four histone core proteins: H2A, H2B, H3, and H4 on rBMSCs. These changes represent novel insights into the aging process and could have implications regarding the potential for autologous stem cells therapy in older patients. 相似文献
5.
Kinsuk Das Bhabesh Mili Madhusoodan A.P. Abhishek Chandra Saxena Ajay Kumar Praveen Singh Med Ram Verma Mihir Sarkar Sadhan Bag 《Tissue & cell》2017,49(2):270-274
Stem cell niche research uses nanotechnologies to mimic the extra-cellular microenvironment to promote proliferation and differentiation. The aim of designing different scaffolds is to simulate the best structural and environmental pattern for extracellular matrix. This experiment was designed to study the proliferative behaviour of canine bone marrow deriver mesenchymal stem cells (MSCs) on different nanomaterial based thin film scaffolds of carbon nanotubes (CNT), chitosan and poly ε-caprolactone. Similar number of cells was seeded on the scaffolds and standard cell culture flask, taken as control. Cells were maintained on DMEM media and relative number of metabolically active cells was determined by MTT assay up to day six of culture. Cells proliferated on control and all the scaffolds as the days progressed. Although proliferation rate was slow but no decline of cell number was noticed on the scaffolds during the study period. Initially, the cell proliferation was lower on CNT but as time progressed no significant difference was observed compared to control. The result indicated that nanomaterial based scaffolds reduce the proliferation rate of canine MSCs. However, canine MSCs adapted and proliferated better on CNT substrate in vitro and may be used as a scaffold component in canine tissue engineering in future. 相似文献
6.
Jin Zhou Ye Zhang Qiuxia Lin Zhiqiang Liu Haibin Wang Cuimi Duan Yanmeng Wang Tong Hao Kuiwu Wu Changyong Wang 《遗传学报》2010,37(7):451-460
Embryonic stem (ES) cells have the potential to develop into any type of tissue and are considered as a promising source of seeding cells for tissue engineering and transplantation therapy. The main catalyst for ES cells differentiation is the growth into embryoid bodies (EBs), which are utilized widely as the trigger of in vitro differentiation. In this study, a novel method for generating EBs from mouse ES cells through culture in collagen/Matrigel scaffolds was successfully established. When single ES cells were seeded in three dimensional collagen/Matrigel scaffolds, they grew into aggregates gradually and formed simple EBs with circular structures. After 7 days' culture,they formed into cystic EBs that would eventually differentiate into the three embryonic germ layers. Evaluation of the EBs in terms of morphology and potential to differentiate indicated that they were typical in structure and could generate various cell types; they were also able to form into tissue-like structures. Moreover, with introduction of ascorbic acid, ES cells differentiated into cardiomyocytes efficiently and started contracting synchronously at day 19. The results demonstrated that collagen/Matrigel scaffolds supported EBs formarion and their subsequent differentiation in a single three dimensional environment. 相似文献
7.
In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel 总被引:2,自引:0,他引:2
Yokoyama A Sekiya I Miyazaki K Ichinose S Hata Y Muneta T 《Cell and tissue research》2005,322(2):289-298
Graft implantation is one of the more popular procedures for repairing cartilage defects; however, sacrifices of the donor
site have been an issue. Mesenchymal stem cells (MSCs) are a fascinating source for regenerative medicine because they can
be harvested in a less invasive manner and are easily isolated and expanded, with multipotentiality including chondrogenesis.
MSCs can be isolated from various adult mesenchymal tissues including synovium. Here, we attempted to form cartilage from
the composites of synovium-derived MSCs with collagen gel in vitro. After 21 days of culture, the composites had increased
their cartilage matrix, as demonstrated by toluidine blue staining and immunohistochemistry for type II collagen. The composites
consisting of 5×107 and 108 cells/ml in gel were richer in proteoglycans than those consisting of lower cell densities. After 1 day, MSCs/gel composites
contracted and the diameter decreased by 30%; however, they were stable thereafter. Round cells with short processes producing
collagen fibrils showing a similar morphology to that of chondrocytes were seen in the composites by transmission electron
microscopy. During composite culture, chondroitin sulfate and mRNA expression for cartilage-related genes increased, demonstrating
cartilage maturation. Using an optimized method, we obtained cartilage discs with a diameter of 7 mm and a thickness of 500 μm.
Our procedure should thus make it possible to produce a large cartilage matrix in vitro. The tissue engineering of autologous
cartilage from the composites of synovium-derived MSCs with collagen gel in vitro for transplantation may be a future alternative
to graft implantation for patients with cartilage defects.
This study is supported in part by grants from the Japanese Society for the Promotion of Science (16591478), the Japanese
Orthopaedics and Traumatology Foundation, and the Nakatomi Foundation to I.S., and the Japanese Society for the Promotion
of Science (16591477), the Japanese Sports Medicine Foundation, the Japanese Latest Osteoarthritis Society, and the Center
of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone in Tokyo Medical
and Dental University to T.M. 相似文献
8.
Optimization of oocyte culture conditions is a crucial aspect of reproductive biology and technology. In the present study, maturation of germinal vesicle-stage marmoset oocytes were evaluated in the following media: Waymouth medium, Waymouth medium containing porcine follicular fluid (pFF) (Waymouth-pFF medium), and porcine oocyte medium (POM). Oocytes cultured in Waymouth-pFF medium had higher maturation rates to the metaphase II stage than those cultured in Waymouth medium (36.1% vs. 24.8%, respectively, P < 0.05), indicating the suitability of this medium for culturing marmoset oocytes. Hence, maturation of marmoset oocytes cultured in POM was subsequently evaluated. The rate of maturation to the metaphase I stage was significantly higher and degradation rates were significantly lower in oocytes cultured in POM than those cultured in Waymouth medium. In addition, three offspring were successfully obtained after transfer of embryos matured in chemically defined medium. Therefore, we concluded that POM was suitable for marmoset oocyte culture. Furthermore, this was apparently the first report of marmoset offspring derived from oocytes cultured in chemically defined medium. 相似文献
9.
Callitrichids are cooperative breeders, characterized by obligate twinning, extensive paternal care, and monopolization of reproduction by the dominant female. This is the case in the common marmoset, and in common marmoset groups of more than one adult female, subordinate females are typically acyclic consistent with infertility. However, one callitrichid, Goeldi's monkey, gives birth to singletons and exhibits low paternal care. Given these reproductive traits of Goeldi's monkey, we hypothesized that there would not be suppression of ovarian activity. To test this hypothesis, we applied non-invasive endocrine methods in a step-wise experiment with laboratory groups of both species. In each species, six pairs of sisters were studied alone, in visual contact with an unrelated male and in a polygynous trio with the male, and urine samples were collected for determination of oestrogen titres reflecting ovarian activity. Common marmoset sister pairs exhibited a marked difference in social status: during the study 5 of 6 dominant females conceived but only 1 of 6 subordinate females; the remaining 5 subordinates were acyclic at the end of the study, and instances of ovulation typically resulted in aggression. Goeldi's monkey sister pairs showed no status differences: in all pairs, however, both sisters exhibited a temporary cessation of ovarian cyclicity on trio formation, followed by ovulation and conception. We conclude that these marked differences in ovarian responses reflect the differences in inter-female competition for paternal caregiving resources. In common marmosets with high inter-female competition, suppression of ovulation functions to reduce aggression received by subordinate females; in Goeldi's monkey with low competition, temporary cessation of ovulation could facilitate female choice. 相似文献
10.
11.
Adipose tissue is a major metabolic organ, and it has been traditionally classified as either white adipose tissue(WAT) or brown adipose tissue(BAT). WAT and BAT are characterized by different anatomical locations, morphological structures, functions, and regulations. WAT and BAT are both involved in energy balance. WAT is mainly involved in the storage and mobilization of energy in the form of triglycerides, whereas BAT specializes in dissipating energy as heat during cold- or diet-induced thermogenesis. Recently, brownlike adipocytes were discovered in WAT. These brownlike adipocytes that appear in WAT are called beige or brite adipocytes. Interestingly, these beige/brite cells resemble white fat cells in the basal state, but they respond to thermogenic stimuli with increased levels of thermogenic genes and increased respiration rates. In addition, beige/brite cells have a gene expressionpattern distinct from that of either white or brown fat cells. The current epidemic of obesity has increased the interest in studying adipocyte formation(adipogenesis), especially in beige/brite cells. This review summarizes the developmental process of adipose tissues that originate from the mesenchymal stem cells and the features of these three different types of adipocytes. 相似文献
12.
目的:研究低温冻存对兔脂肪间充质干细胞部分生物学特性的影响。方法采用组织块法分离培养兔脂肪间充质干细胞。用倒置显微镜观察原代细胞的细胞形态,流式细胞仪检测兔脂肪间充质干细胞的免疫表型。取第3代兔脂肪间充质干细胞置于-196℃液氮保存半年,37℃复苏并传至第7代。实验分为两组,实验组为冻存复苏后传至第7代的兔脂肪间充质干细胞,对照组为未冻存的第7代兔脂肪间充质干细胞,用MTT绘制其生长曲线;添加成脂、成骨诱导液进行诱导,油红O、茜素红染色和碱性磷酸酶活性检测分别进行鉴定。结果体外培养的兔脂肪间充质干细胞呈梭形纤维样细胞形态,生长力旺盛。流式细胞仪检测显示,第3代兔脂肪间充质干细胞强表达CD44、CD90,阴性表达造血细胞相关的表面标志CD45。两组细胞生长曲线呈典型的“S”形,无统计学差异(P>0.05);成脂诱导14 d后,油红O染色呈阳性;成骨诱导2周时茜素红染色阳性,ALP表达活性随成骨诱导时间延长不断增加且无统计学差异( P>0.05)。结论冻存后的兔脂肪间充质干细胞体外生长及多向分化潜能未发生显著变化。 相似文献
13.
Bone marrow mesenchymal stem cells (MSCs) can develop into hematopoietic and mesenchymal lineages but have not been known to participate in the production of retina. Here we report that bone marrow mesenchymal stem cells, after being subretinally transplanted into normal or Nd: YAG laser-injured rat eye, can integrate into RPE layer, photoreceptor layer, bipolar cell layer and ganglion layer. DAPI-labeling detection was used to trace the origin of the repopulating cells. DAPI fluorescence was used to identify retina cells of bone marrow origin 10, 20, 35 and 50 days after transplantation. No formation of rosettes was found but some random cells were found at the end of the observation. MSCs-originated cells spread more widely in the injured retinas than in the normal ones. Immunohistochemical detection showed that though the cells could express neuronal nuclei (NeuN), neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and cytokeratin (CK), the proteins expression in the injured transplantation group was abnormal in some region compared with that in the normal transplantation group. Electroretinogram (ERG) showed that ERG-b wave of the injured transplantation group is significantly higher than that of the two laser-injured control groups. These results suggest that a proportion of MSCs can differentiate into retina-like structure in vivo and the differentiation differs in normal and laser-injured retinas. 相似文献
14.
Xinghui Song Yanwei Li Xiao Chen Guoli Yin Qiong Huang Yingying Chen Guowei Xu Linlin Wang 《Genetics and molecular biology》2014,37(1):127-134
In this work we describe the establishment of mesenchymal stem cells (MSCs) derived from embryonic stem cells (ESCs) and the role of bFGF in adipocyte differentiation. The totipotency of ESCs and MSCs was assessed by immunofluorescence staining and RT-PCR of totipotency factors. MSCs were successfully used to induce osteoblasts, chondrocytes and adipocytes. MSCs that differentiated into adipocytes were stimulated with and without bFGF. The OD/DNA (optical density/content of total DNA) and expression levels of the specific adipocyte genes PPARγ2 (peroxisome proliferator activated receptor γ2) and C/EBPs were higher in bFGF cells. Embryonic bodies had a higher adipocyte level compared with cells cultured in plates. These findings indicate that bFGF promotes adipocyte differentiation. MSCs may be useful cells for seeding in tissue engineering and have enormous therapeutic potential for adipose tissue engineering. 相似文献
15.
R. Gonzalez V. Vargas P. SantaCruz M. Schiewe A. Patel 《Biochemical and biophysical research communications》2009,385(4):570-1752
Mesenchymal stem cells (MSCs) isolated from several adult human tissues are reported to be a promising tool for regenerative medicine. In order to broaden the array of tools for therapeutic application, we isolated a new population of cells from adult human testis termed gonadal stem cells (GSCs). GSCs express CD105, CD166, CD73, CD90, STRO-1 and lack hematopoietic markers CD34, CD45, and HLA-DR which are characteristic identifiers of MSCs. In addition, GSCs express pluripotent markers Oct4, Nanog, and SSEA-4. GSCs propagated for at least 64 population doublings and exhibited clonogenic capability. GSCs have a broad plasticity and the potential to differentiate into adipogenic, osteogenic, and chondrogenic cells. These studies demonstrate that GSCs are easily obtainable stem cells, have growth kinetics and marker expression similar to MSCs, and differentiate into mesodermal lineage cells. Therefore, GSCs may be a valuable tool for therapeutic applications. 相似文献
16.
Hui Ke Peng Wang Weihua Yu Xiaoming Liu Chang Liu Fan Yang Frank Fuxiang Mao Liangming Zhang Xiuming Zhang Bruce T. Lahn Andy Peng Xiang 《Differentiation; research in biological diversity》2009
Mesenchymal stem cells (MSCs) have received considerable attention in recent years. Particularly exciting is the prospect that MSCs could be differentiated into specialized cells of interest, which could then be used for cell therapy and tissue engineering. MSCs derived from nonhuman primates could be a powerful tool for investigating the differentiation potential in vitro and in vivo for preclinical research. The purpose of this study was to isolate cynomolgus mesenchymal stem cells (cMSCs) from adult bone marrow and characterize their growth properties and multipotency. Mononuclear cells were isolated from cynomolgus monkey bone marrow by density-gradient centrifugation, and adherent fibroblast-like cells grew well in the complete growth medium with 10 μM Tenofovir. cMSCs expressed mesenchymal markers, such as CD29, CD105, CD166 and were negative for hematopoietic markers such as CD34, CD45. Furthermore, the cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages under certain conditions, maintaining normal karyotype throughout extended culture. We also compared different methods (lipofection, nucleofection and lentivirus) for genetic modification of cMSCs and found lentivirus proved to be the most effective method with transduction efficiency of up to 44.6% and lowest level of cell death. The cells after transduction stably expressed green fluorescence protein (GFP) and maintained the abilities to differentiate down osteogenic and adipogenic lineages. In conclusion, these data showed that cMSCs isolated from cynomolgus bone marrow shared similar characteristics with human MSCs and might provide an attractive cell type for cell-based therapy in higher-order mammalian species disorder models. 相似文献
17.
Makoto Moro Ryuzo Torii Hitoshi Koizumi Yoichi Inada Yasuo Etoh Hiroshi Miyata Yoshikuni Tanioka 《Primates; journal of primatology》1995,36(2):249-257
A homologous double-antibody radioimmunoassay developed for humans was used to measure serum prolactin, progesterone, and
estradiol in common marmosets. In the ovarian cycle of common marmosets, serum progesterone began to increase after an estradiol
surge, attained a peak level, and then declined before the ensuing pre-ovulatory rise in estradiol. During the luteal phase,
the change in serum concentrations of estradiol was synchronized with that of progesterone. During the ovarian cycle there
was no consistent change in serum prolactin concentrations. During the last 75 days of pregnancy the prolactin level was higher
than during the ovarian cycle and the first 70 days of pregnancy. Moreover, during lactation, mothers with suckling twin infants
had a higher prolactin level than during the final stage of pregnancy. 相似文献
18.
Laparoscopic observations of morphological changes of the ovary during the ovarian cycle in conjunction with radioimmunoassay
of serum progesterone and estradiol-17β was investigated as a method of monitoring the ovarian cycle in the common marmoset.
In the common marmoset, plural follicles first appeared in each ovary five days prior to ovulation. At three to four days
prior to ovulation one or two follicles developed into translucent blisters on the surface of the ovary. As the follicles
filled with follicular fluid, they became larger and clearer until one to two days prior to ovulation, at which time they
formed well defined, transparent bubbles protruding from the surface of the ovary. After ovulation, the ovulation point could
be detected at the center of the follicle, sometimes surrounded by a corpus of engorged blood vessels. Ovulations of the plural
follicles were not simultaneous, and due to the time lag ovulations took at least 12 to 20 hrs in four out of seven animals
examined. After two to five days of ovulation the corpus hemorragicum, a bright red protrusion made of tissue and blood disrupted
by ovulation, was found. Subsequently, the color of the formatted corpus luteum changed from dark-red to yellow then to yellow
white. While the corpus luteum remained reddish in color serum progesterone was maintained at as high levels as in the luteal
phase.
There was no mature follicle or corpus luteum in subordinate female ovaries. 相似文献
19.
20.
Ladda Meesuk Chairat Tantrawatpan Pakpoom Kheolamai Sirikul Manochantr 《Biochemistry and Biophysics Reports》2016
Mesenchymal stromal cells derived from amnion (AM-MSCs) can be easily obtained in large quantity by less invasive method in comparison to bone marrow-derived MSCs (BM-MSCs). However, the biological and immunosuppressive properties of AM-MSCs are still poorly characterized. Previous studies demonstrated that BM-MSCs expressed indoleamine 2,3-dioxygenase (IDO) to suppress T-cell responses. This study was designed to address whether IDO contributes to the immunosuppressive function of AM-MSCs. MSCs isolated from amnion were cultured in complete medium similar to BM-MSCs. After culture, AM-MSCs exhibited spindle shape morphology and expressed MSC markers similar to that of BM-MSCs. In addition, AM-MSCs were able to differentiate into adipocytes and osteoblasts. Fascinatingly, AM-MSCs and BM-MSCs exhibited comparable degree of immunosuppressive effect when they were co-cultured with activated T-cells. In addition, IDO secreted by AM-MSCs was responsible for induction of immunosuppressive activities in the same manner as BM-MSCs. Taken together; the results of the present study demonstrate that while AM-MSCs and BM-MSCs show similar immunosuppressive effect, AM-MSCs may have additional advantage over the BM-MSCs in terms of availability. Therefore, AM-MSCs might be considered a potential source for therapeutic applications especially for treatment of immune related diseases. 相似文献