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21.
Expansion strategies for human mesenchymal stromal cells culture under xeno‐free conditions 下载免费PDF全文
Patrícia Aparecida Tozetti Samia Rigotto Caruso Amanda Mizukami Taisa Risque Fernandes Fernanda Borges da Silva Fabiola Traina Dimas Tadeu Covas Maristela Delgado Orellana Kamilla Swiech 《Biotechnology progress》2017,33(5):1358-1367
Choosing the culture system and culture medium used to produce cells are key steps toward a safe, scalable, and cost‐effective expansion bioprocess for cell therapy purposes. The use of AB human serum (AB HS) as an alternative xeno‐free supplement for mesenchymal stromal cells (MSC) cultivation has increasingly gained relevance due to safety and efficiency aspects. Here we have evaluated different scalable culture systems to produce a meaningful number of umbilical cord matrix‐derived MSC (UCM MSC) using AB HS for culture medium supplementation during expansion and cryopreservation to enable a xeno‐free bioprocess. UCM MSC were cultured in a scalable planar (compact 10‐layer flasks and roller bottles) and 3‐D microcarrier‐based culture systems (spinner flasks and stirred tank bioreactor). Ten layer flasks and roller bottles enabled the production of 2.6 ± 0.6 × 104 and 1.4 ± 0.3 × 104 cells/cm2. UCM MSC‐based microcarrier expansion in the stirred conditions has enabled the production of higher cell densities (5.5–23.0 × 104 cells/cm2) when compared to planar systems. Nevertheless, due to the moderate harvesting efficiency attained, (80% for spinner flasks and 46.6% for bioreactor) the total cell number recovered was lower than expected. Cells maintained the functional properties after expansion in all the culture systems evaluated. The cryopreservation of cells (using AB HS) was also successfully carried out. Establishing scalable xeno‐free expansion processes represents an important step toward a GMP compliant large‐scale production platform for MSC‐based clinical applications. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1358–1367, 2017 相似文献
22.
Maud Gits-Muselli Marie-Noelle Peraldi Nathalie de Castro Véronique Delcey Jean Menotti Nicolas Guigue Samia Hamane Emmanuel Raffoux Anne Bergeron Sandrine Valade Jean-Michel Molina Stéphane Bretagne Alexandre Alanio 《PloS one》2015,10(5)
Pneumocystis pneumonia is a severe opportunistic infection in immunocompromised patients caused by the unusual fungus Pneumocystis jirovecii. Transmission is airborne, with both immunocompromised and immunocompetent individuals acting as a reservoir for the fungus. Numerous reports of outbreaks in renal transplant units demonstrate the need for valid genotyping methods to detect transmission of a given genotype. Here, we developed a short tandem repeat (STR)-based molecular typing method for P. jirovecii. We analyzed the P. jirovecii genome and selected six genomic STR markers located on different contigs of the genome. We then tested these markers in 106 P. jirovecii PCR-positive respiratory samples collected between October 2010 and November 2013 from 91 patients with various underlying medical conditions. Unique (one allele per marker) and multiple (more than one allele per marker) genotypes were observed in 34 (32%) and 72 (68%) samples, respectively. A genotype could be assigned to 55 samples (54 patients) and 61 different genotypes were identified in total with a discriminatory power of 0.992. Analysis of the allelic distribution of the six markers and minimum spanning tree analysis of the 61 genotypes identified a specific genotype (Gt21) in our hospital, which may have been transmitted between 10 patients including six renal transplant recipients. Our STR-based molecular typing method is a quick, cheap and reliable approach to genotype Pneumocystis jirovecii in hospital settings and is sensitive enough to detect minor genotypes, thus enabling the study of the transmission and pathophysiology of Pneumocystis pneumonia. 相似文献
23.
Samia Dabbou Imed Rjiba Adel Echbili Noureddine Gazzah Beligh Mechri Mohamed Hammami 《化学与生物多样性》2010,7(7):1801-1813
Our study reports the triglyceride (triacylglycerol, TAG) composition of five new Tunisian virgin olive oil cultivars obtained through controlled crossings of the cultivar (cv.) Chemlali Sfax. These cultivars have been selected among a progeny of 500 olive descendants, based on an evaluation of the fatty‐acid (FA) composition of their oils. Among these samples, two were derived from the crossing with the cv. Sigoise as pollinator (SM634) or pollen acceptor (SM1110) and the others from the crossing with the cv. Meski as pollen acceptor (SM513, SM514, and SM517). The five descendants were characterized by a good fat value, a balanced FA composition, and a high content of triolein, varying between 26.9 (SM514) and 45.46% (SM1110). They had an improved FA composition as compared to that of the cv. Chemlali Sfax and their fruits were slightly bigger. The principal component analysis suggested that the TAG variables were more suitable than the total FAs for an optimum classification of the cultivar samples analyzed. The cultivars obtained through the crossing with the cv. Sigoise (as pollen acceptor or pollinator) had a more favorable composition of FAs and TAG than those obtained through the crossing with the cv. Meski, which indicated that genetic factors had the most important influence on the quality of the virgin olive oils. 相似文献
24.
Li Y Laue K Temtamy S Aglan M Kotan LD Yigit G Canan H Pawlik B Nürnberg G Wakeling EL Quarrell OW Baessmann I Lanktree MB Yilmaz M Hegele RA Amr K May KW Nürnberg P Topaloglu AK Hammerschmidt M Wollnik B 《American journal of human genetics》2010,87(6):757-767
Altered Bone Morphogenetic Protein (BMP) signaling leads to multiple developmental defects, including brachydactyly and deafness. Here we identify chondroitin synthase 1 (CHSY1) as a potential mediator of BMP effects. We show that loss of human CHSY1 function causes autosomal-recessive Temtamy preaxial brachydactyly syndrome (TPBS), mainly characterized by limb malformations, short stature, and hearing loss. After mapping the TPBS locus to chromosome 15q26-qterm, we identified causative mutations in five consanguineous TPBS families. In zebrafish, antisense-mediated chsy1 knockdown causes defects in multiple developmental processes, some of which are likely to also be causative in the etiology of TPBS. In the inner ears of zebrafish larvae, chsy1 is expressed similarly to the BMP inhibitor dan and in a complementary fashion to bmp2b. Furthermore, unrestricted Bmp2b signaling or loss of Dan activity leads to reduced chsy1 expression and, during epithelial morphogenesis, defects similar to those that occur upon Chsy1 inactivation, indicating that Bmp signaling affects inner-ear development by repressing chsy1. In addition, we obtained strikingly similar zebrafish phenotypes after chsy1 overexpression, which might explain why, in humans, brachydactyly can be caused by mutations leading either to loss or to gain of BMP signaling. 相似文献
25.
Abstract. Telomerase is a ribonucleoprotein that can maintain telomeres, the repetitive sequences of DNA found at the end of eukaryotic chromosomes, and confer long-term proliferative capacity on cells. Telomerase expression is essential during periods of intense cell division such as the early developmental process. In later development, some species retain telomerase activity while others repress telomerase activity in what is thought to be a tumor-protective mechanism. Despite the importance of telomerase expression in both development and neoplastic disease, no studies to date have characterized its expression in bivalves. We present the first report of telomerase expression in a bivalve species, the sand scallop, Euvola ziczac. Telomerase activity was detected throughout the early stages of development and in all adult tissues examined. Analysis of DNA isolated from adult tissues indicated long telomeres, with terminal restriction fragment lengths >20 kb in both somatic and germ tissues. Ubiquitous telomerase expression throughout development and into adulthood would suggest a lack of telomere-related senescence and suggests that these scallops do not use telomerase repression as a mechanism to suppress the formation of neoplasm. 相似文献
26.
Development of a measles vaccine production process in MRC-5 cells grown on Cytodex1 microcarriers and in a stirred bioreactor 总被引:1,自引:0,他引:1
Measles vaccination remains the most efficient way to control the spread of the virus. This work focuses on the production
of a measles vaccine using stirred conditions as an advanced option for process scale up. Non-porous Cytodex 1 microcarriers
were used to support MRC-5 cell growth in suspension cultures. Virus replication was first optimized in spinner flasks, and
the effects of various operational parameters were investigated. Cell infection with AIK-C measles strain at an MOI (multiplicity
of infection) of 0.005, without glucose regulation and in M199 medium, resulted in a virus titer of 106.25 TCID50 (median tissue culture infective dose)/ml. To optimize the production process in a 7-l bioreactor, we carried out various
perfused cultures using minimum essential medium (MEM) + 5% FCS diluted with phosphate-buffered saline (PBS). We achieved
a high cell density level (4.1 × 106 cells/ml) with an efficient use of the medium when MEM + 5% FCS diluted with PBS at 25% was used during the cell amplification
step. Optimization of measles production in MRC-5 cells grown on Cytodex 1 beads in a 7-l bioreactor showed that perfusion
was the most efficient when compared to repeated-batch culture. Perfusion at a rate of 0.25 V (reactor volume)/day showed
the highest specific productivity (1.6 IVP [infectious virus particle] cell−1 day−1). Testing of several stabilizers containing pharmaceutically improved components such as sugars, amino acids, and charged
ions showed that the formulation composed of sucrose and MgCl2, led to the maintenance of the infectivity of the AIK-C measles virus strain to a significant level, when stored at +28 °C,
+4 °C and −60 °C. 相似文献
27.
Effect of acyl donor chain length and sugar/acyl donor molar ratio on enzymatic synthesis of fatty acid fructose esters 总被引:3,自引:0,他引:3
Samia Soultani Jean-Marc Engasser Mohamed Ghoul 《Journal of Molecular Catalysis .B, Enzymatic》2001,11(4-6):725-731
Lipase-catalyzed synthesis of fatty acid sugar esters through direct esterification was performed in 2-methyl 2-butanol as solvent. Fructose and saturated fatty acids were used as substrates and the reaction was catalyzed by immobilized Candida antarctica lipase. The effect of the initial fructose/acyl donor molar ratio and the carbon-chain length of the acyl donor as well as their reciprocal interactions on the reaction performance were investigated. For this purpose, an experimental design taking into account variations of the molar ratio (from 1:1 to 1:5) and the carbon-chain length of the fatty acid (from C8 to C18) was employed. Statistical analysis of the data indicated that the two factors as well as their interactions had significant effects on the sugar esters synthesis. The obtained results showed that whatever the molar ratio used, the highest concentration (73 g l−1), fructose and fatty acid conversion yields (100% and 80%, respectively) and initial reaction rate (40 g l−1 h−1) were reached when using the C18 fatty acid as acyl donor. Low molar ratios gave the best fatty acid conversion yields and initial reaction rates, whereas the best total sugar ester concentrations and fructose conversion yields were obtained for high molar ratios. 相似文献
28.
Dan Hanson Philip G. Murray Amit Sud Samia A. Temtamy Andrea Superti-Furga Jill Urquhart Emma Hilton Peter Scambler Peter E. Clayton 《American journal of human genetics》2009,84(6):801-806
3-M syndrome is an autosomal-recessive primordial growth disorder characterized by significant intrauterine and postnatal growth restriction. Mutations in the CUL7 gene are known to cause 3-M syndrome. In 3-M syndrome patients that do not carry CUL7 mutations, we performed high-density genome-wide SNP mapping to identify a second locus at 2q35-q36.1. Further haplotype analysis revealed a 1.29 Mb interval in which the underlying gene is located and we subsequently discovered seven distinct null mutations from 10 families within the gene OBSL1. OBSL1 is a putative cytoskeletal adaptor protein that localizes to the nuclear envelope. We were also able to demonstrate that loss of OBSL1 leads to downregulation of CUL7, implying a role for OBSL1 in the maintenance of CUL7 protein levels and suggesting that both proteins are involved within the same molecular pathway. 相似文献
29.
The interconnection network is one of the key architectural components in any parallel computer. The distribution of the traffic
injected into the network is among the factors that greatly influences network performance. The uniform traffic pattern has
been adopted in many existing network performance evaluation studies due to the tractability of the resulting analytical modelling
approach. However, many real applications exhibit non-uniform traffic patterns such as hot-spot traffic. K-ary n-cubes have been the mostly widely used in the implementation of practical parallel systems. Extensive research studies have
been conducted on the performance modelling and evaluation of these networks. Nonetheless, most of these studies have been
confined to uniform traffic distributions and have been based on software simulation. The present paper proposes a new stochastic
model to predict message latency in k-ary
n-cubes with deterministic routing in the presence of hot-spot traffic. The model has been validated through simulation experiments
and has shown a close agreement with simulation results.
相似文献
Geyong MinEmail: |
30.