Abstract Mesenchymal stem/stromal cells (MSCs) are multipotent somatic stem/progenitor cells that can be isolated from various tissues and have attracted increasing attention from the scientific community. This is due to MSCs showing great potential for incurable disease treatment, and most applications of MSCs involve tissue degeneration and treatment of immune- and inflammation-mediated diseases. Conventional MSC cultures contain fetal bovine serum (FBS), which is a common supplement for cell development but is also a risk factor for exposure to animal-derived pathogens. To avoid the risks resulting from the xenogeneic origin and animal-derived pathogens of FBS, xeno-free media have been developed and commercialized to satisfy MSC expansion demands for human clinical applications. This review summarized and provided an overview of xeno-free media that are currently used for MSC expansion. Additionally, we discussed the influences of different xeno-free media on MSC biology with particular regard to cell morphology, surface marker expression, proliferation, differentiation and immunomodulation. The xeno-free media can be serum-free and xeno-free media or media supplemented with some human-originating substances, such as human serum, human platelet lysates, human umbilical cord serum/plasma, or human plasma-derived supplements for cell culture medium. These media have capacity to maintain a spindle-shaped morphology, the expression of typical surface markers, and the capacity of multipotent differentiation and immunomodulation of MSCs. Xeno-free media showed potential for safe use for human clinical treatment. However, the influences of these xeno-free media on MSCs are various and any xeno-free medium should be examined prior to being used for MSC cultures.

Abstract Mesenchymal stem/stromal cells (MSCs) are multipotent somatic stem/progenitor cells that can be isolated from various tissues and have attracted increasing attention from the scientific community. This is due to MSCs showing great potential for incurable disease treatment, and most applications of MSCs involve tissue degeneration and treatment of immune- and inflammation-mediated diseases. Conventional MSC cultures contain fetal bovine serum (FBS), which is a common supplement for cell development but is also a risk factor for exposure to animal-derived pathogens. To avoid the risks resulting from the xenogeneic origin and animal-derived pathogens of FBS, xeno-free media have been developed and commercialized to satisfy MSC expansion demands for human clinical applications. This review summarized and provided an overview of xeno-free media that are currently used for MSC expansion. Additionally, we discussed the influences of different xeno-free media on MSC biology with particular regard to cell morphology, surface marker expression, proliferation, differentiation and immunomodulation. The xeno-free media can be serum-free and xeno-free media or media supplemented with some human-originating substances, such as human serum, human platelet lysates, human umbilical cord serum/plasma, or human plasma-derived supplements for cell culture medium. These media have capacity to maintain a spindle-shaped morphology, the expression of typical surface markers, and the capacity of multipotent differentiation and immunomodulation of MSCs. Xeno-free media showed potential for safe use for human clinical treatment. However, the influences of these xeno-free media on MSCs are various and any xeno-free medium should be examined prior to being used for MSC cultures.

Objective: To examine the multiplication efficiency Japanese encephalitis virus (JEV) genotype I (GI) and genotype III (GIII) of different cell lines which originated from human, porcine, mosquitoes in order to prove mechanism of JEV GI replacement JEV GIII since it emerging in nature recent decades. Methods: The mixture of GI and GIII JEV isolates was inoculated on human rhabdomyosarcoma (RD), pig kidney epithelial (PS) and Aedes albopictus C6/36 clone (C6/36) which originated from human, porcine and mosquitoes, respectively. Plaque assays were performed to calculate virus titer and real-time RT-PCR with GI and GIII specific primer sets to quantify the number of GI and GIII RNA copies. Results: The highest virus titer reached at the 3rd day of post infection when GI and GIII mixture was inoculated on RD and PS and that of C6/36 was at the 4th day. JEVs were amplified and maintained by C6/36 cells after 10 passages whereas that by RD and PS only limited within 8 and 6 passages, respectively. GI strain amplified and maintained more efficiently on C6/36 and PS but not RD, whereas GIII strain amplified and maintained more efficiently on RD. Conclusions: There is a correlation between the multiplication efficiency of GI and GIII JEV strains when these two genotype strains co-infected on different cell lines with the predominance of GI strains in C6/36 and PS and the limited detection of GI strains in RD cells proving a possible mechanism of shift JEV genotypes in nature recent decades since GI emerging.

Objective: To establish a surveillance in Dong Thap, at the border with Cambodia by assessing the presence of DENV serotypes and CHIKV among patients hospitalized at Dong Thap general hospital. Methods: Cross-sectional descriptive analysis was conducted on a cohort of 131 patients hospitalized with acute fever and symptoms compatible with dengue or chikungunya. The study was conducted from January 2012 to February 2013. The full clinical picture was established as well as serological and molecular detection. Serological analysis was sequentially performed on blood samples collected on admission and an average of seven days after admission. The detection of IgM antibody to DENV was performed by IgM capture ELISA and the detection of DENV and CHIKV RNA was done by reverse-transcription multiplex PCR. Results: 101 patients out of 131 (77%) were confirmed with dengue. All four dengue serotypes were detected with a predominance of DENV2 and DENV4. No chikungunya infection was detected although reported in neighboring Cambodia. A differential efficiency of serological dengue detection was observed. Efficiency was 29% upon admission and 53% after seven days on the same patients. 30 patients out of 131 (23%) were negative with both DENV and CHIKV. Conclusions: Dengue is at risk of being underestimated and chikungunya is not systematically detected. Changes in detection and surveillance procedures are therefore discussed to increase efficiency of dengue detection and continue the monitoring the emergence of CHIKV in Dong Thap province and in Vietnam.

Background: Recently Japanese Encephalitis (JE) virus type 1 has surfaced and is co-circulated with JE virus type 3 in the northern areas of Viet Nam, so a sensitivity of JE viral antigen genotype 3 to detect IgM is required. Objectives: To compare the sensitivity of JE viral antigen genotype 1 and 3 to detect IgM against the JE virus. Materials and method: 783 cerebrospinal fluid and serum samples from viral encephalitis cases from 1999-2005 were collected and examined by MAC-ELISA for JE viral antigen genotype 1 and 3. Results: The agreement on the diagnosis of these kinds of antigen was 99.7% and the sensitivity of JE viral antigen genotype 3 was higher than that of genotype 1. Thus, JE viral antigen genotype 3 could be considered as the selected antigen for JE diagnosis in Viet Nam. IgM titer determined by JE viral antigen genotype 1 was higher than that of genotype 3 in 2003 and 2005 and lower in 1999, 2000, 2001, 2002 and 2004. Conclusion: The dominant phenomenon of JE viral genotypes differing over the years might be due to the interaction of the virus and its vectors. Further study is required to clarify this observation.
Japanese Encephalitis ; antigen

Background: Mosquitoes and pigs play important roles in maintaining and increasing the Japanese Encephalitis (JE) virus in nature and which is then transmitted to humans. Thus, surveillance of the JE infection frequency in the pig population may predict the human JE cases. \r\n', u'Objectives: The study aimed to determine IgG antibody against the JE virus in the pig population in Hanam province \r\n', u'Subjects and methods: The study included 1791 pig serum samples collected from 3 districts of Hanam province from Apr 2006 to Mar 2007. GAC-ELISA technique was used to determine the JE virus infection in the swine population.\r\n', u'Results: The average positive rate in pig population was 34.9 % (626/1791); with the highest frequency occurring in the summer (37.7%- 84.0 %), co-incident with the JE season in Northern Vietnam. On the contrary, in winter JE case are rare, frequency of IgG antibody against JE virus in the swine population was low, ranging from 9.2% to 22.0.%. \r\n', u'Conclusions: These results have shown the ecologically close relationship between the amplification of the JE virus in the swine population, vector and JE cases in northern Vietnam. \r\n', u'
Japanese encephalitis ; pig population ; GAC-ELISA.

Background: Japanese Encephalitis (JE) virus is a leading cause of encephalitis in children with high mortality and complication. JE is a dangerous infectious disease via Culex mosquitoes. Objective: To identify the density of some mosquitoes and vectors that transmit JE virus in Ha Nam province. Subject and Method:There were 30.333 mosquitoes including 05 genera and 15 species which were collected at night from April 2006 to March 2007 in the following communes: Tanson (midland area) Hungcong (plain area) and Liemchinh (suburban area). Results: A total of 06 Culex species, the highest rate is Culex tritaeniohychus (41.20%); Culex annulus (15.56%); other Culex species occupied around 0.15% -2.49%. In these communes, the density of Culex tritaeniohychus was higher than Culex annulus. They were found to be active all year round, but the highest density of Culex tritaeniohychus and Culex annulus is 5.86 and 2.15, respectively in April and fall to 4.35% and 1.71% in July. The collected mosquitoes during April, May, June and July includes 133 and 28 Culex tritaeniohychus and Culex annulus pools, respectively; all were to be processed for the isolation of Japanese Encephalitis (JE) virus. There were 12 Culex tritaeniohychus and 2 Culex annulus pools which were collected in Tanson commune yielding positive. Conclusion: The results were confirmed that Culex tritaenioohychus and Culex annulus are the major vectors that transmit JE virus in Ha Nam, 2006 - 2007. All 14 JE virus strains were isolated from collected mosquito pools during April, May and June; whereas JE virus strain was not yet isolated from collected mosquito pool in July. Thus, in order to control JE disease, it is necessary to control JE vectors in April, May and June.
Vector ; Culex tritaenioohychus ; Japanese encephalitis ; Ha Nam

Background: IgM antibody capture ELISA (MAC-ELISA) technique has been widely applied for Japanese Encephalitis Virus (JEV) diagnosis. So far rare internationally commercial kits are available. Thus, the international evaluation of the kit is required as per the recommendation of the WHO. Objectives: To evaluate the quality of the IgM antibody capture ELISA diagnostic kit for JEV produced by the Vietnam National Institute of Hygiene and Epidemiology (NIHE). Subjects and method: In this study, NIID kit was used as control to check the kit from NIHE. Both NIHE and NIID kits were used to detect JEV IgM among 38 serum and 6 CFS samples, which belongs to 5 sample groups (JE patients group, dengue patients group, other viral encephalitis patients group, Tick Born Encephalitis (TBE) patient group and healthy JE vaccinated donors group). Results: The detection of JEV IgM by NIHE kit was concurrent with the NIID kit. There is no positive with the JE in the groups of Dengue patients, TBE, other virus encephalitis patients and JE vaccinated donors. Conclusion: MAC-ELISA kit of NIHE can be used for different diagnosis of JEV and Dengue virus (both viruses are in Flavivirus genus), as well as other viruses caused by encephalitis.
IgM antibody ; ELISA diagnostic kit ; Japanese encephalitis virus

Background: In recent year, Japanese Encephalitis Virus (JEV) genotype 1 has been detected among isolates from mosquitoes and pig\u2019s blood samples in northern Viet Nam, but there has been no information on the presence of this genotype in the Central, Southern and Highland regions. Objectives: This study aims to detect the Japanese encephalitis genotype 1 in various different geographic regions of Viet Nam. Material and method: Sequence analysis\u2019s of whole E gene of 18 strains isolated from human, mosquitoes and pig\u2019s blood during 2001-2007. Results: 7 strains isolated from pig\u2019s blood and mosquito samples in the Northern, Central, Southern and Highland fell into genotype 1, but 11 others isolated from humans in the Northern and Central regions belonged to genotype 3. Conclusion: This is the first time that JEV genotype 1 was detected in the central, northern, highland Viet Nam and further studies on genotype 1 causing human diseases needs to be carried out.\r\n', u'\r\n', u'
Virus ; Japanese Encephalitis ; genotype 1 ; E gene.

Background: In recent years, some arbo viruses which causes acute encephalitis syndrome (AES) have been identified in serveral countries in the world such as Chandipura virus belonging to Rhabdoviridae family in India, Banna virus belonging to Reoviridae family in China. In Vietnam, apart from Japanese Encephalitis Virus which is considered as main cause of AES, there are a few intestinal viruses like Herpes symplex virus type 1 and 4 and Nam Dinh virus considering other causes of AES. Objective: To identify the hyppothesis that one virus strain parasitizing in mosquito in Gia Lai province causes AES in order to provide more information about virus strains which cause AES in Vietnam. Subjects and method: Aedes albopictus cell line clone C6/36 was used for the isolation of virus in 43 cerebrospinal fluid samples of patients who were treated in Gia Lai hospital, from January/2005 to July/2005. Result and Conclusion: One virus strain from a 3-year old girl in Gia Lai province was isolated in 2005. The virus coded 05VN225 has the morphology similar to other viruses belonging to Reoviridea family.The nucleic acid sequence of the virus was checked with specific primers of alphavirus and flavirus groups, Nam Dinh virus and Conti virus group B (reovirus) of the Reoviridae. The positive result was confirmed with reovirus primers. This member of the Reoviridae family was isolated from acute encephalitis syndrome in Vietnam in 2005. Further study on pathology of the virus is very necessary.
Encephalitis ; Arbovirus/ epidemiology ; pathology ; Reoviridae/ growth & ; development ; pathogenicity ;