Re-using blood products as an alternative supplement in the optimisation of clinical-grade adipose-derived mesenchymal stem cell culture.

OBJECTIVES: Adipose-derived mesenchymal stem cells (ADMSCs) are a promising strategy for orthopaedic applications, particularly in bone repair. Ex vivo expansion of ADMSCs is required to obtain sufficient cell numbers. Xenogenic supplements should be avoided in order to minimise the risk of infections and immunological reactions. Human platelet lysate and human plasma may be an excellent material source for ADMSC expansion. In the present study, use of blood products after their recommended transfusion date to prepare human platelet lysate (HPL) and human plasma (Hplasma) was evaluated for in vitro culture expansion and osteogenesis of ADMSCs. METHODS: Human ADMSCs were cultured in medium supplemented with HPL, Hplasma and a combination of HPL and Hplasma (HPL+Hplasma). Characteristics of these ADMSCs, including osteogenesis, were evaluated in comparison with those cultured in fetal bovine serum (FBS). RESULTS: HPL and HPL+Hplasma had a significantly greater growth-promoting effect than FBS, while Hplasma exhibited a similar growth-promoting effect to that of FBS. ADMSCs cultured in HPL and/or Hplasma generated more colony-forming unit fibroblasts (CFU-F) than those cultured in FBS. After long-term culture, ADMSCs cultured in HPL and/or Hplasma showed reduced cellular senescence, retained typical cell phenotypes, and retained differentiation capacities into osteogenic and adipogenic lineages. CONCLUSION: HPL and Hplasma prepared from blood products after their recommended transfusion date can be used as an alternative and effective source for large-scale ex vivo expansion of ADMSCs.Cite this article: J. Phetfong, T. Tawonsawatruk, K. Seenprachawong, A. Srisarin, C. Isarankura-Na-Ayudhya, A. Supokawej. Re-using blood products as an alternative supplement in the optimisation of clinical-grade adipose-derived mesenchymal stem cell culture. Bone Joint Res 2017;6:414-422. DOI: 10.1302/2046-3758.67.BJR-2016-0342.R1.

Revelation of staphylococcal cassette chromosome mec types in methicillin-resistant Staphylococcus aureus isolates from Thailand and Vietnam.

Methicillin resistant Staphylococcus aureus (MRSA) is highly prevalent, and its typing plays a crucial role in epidemiology and evolution in both health and community settings. Multiplex PCR and staphylococcal cassette chromosome mec (SCCmec) typing based on mec complexes and cassette chromosome recombinase (ccr) allotypes have been developed for MRSA identification. The first of these procedures can identify 4 mec classes (A, B, C1, and E) and 2 ccr allotypes (B2 and B4) in one tube, and the second can identify mecA, mec class C2, and 3 allotypes (A1, A3, and C). Our method offers a novel means to further differentiate between the main SCCmec types I through XI and is both highly sensitive (detectable up to 0.3ηg DNA) and specific (100%). Several SCCmec types (I, III, IV, V and a non-typeable group) were found in 66 MRSA isolates obtained from Ho Chi Minh City, Vietnam and Nakhon Pathom, Thailand. SCCmec type III was highly predominant in both regions. The designed assay is rapid, convenient, flexible, and reliable. Therefore, this assay is suitable for the high-throughput screening of the main SCCmec types of MRSA isolates.

Evaluation of two screening tests for human leptospirosis.

Leptospirosis is prevalent in Thailand but its diagnosis depends primarily on clinical awareness. Serodiagnosis is of great assistance in the diagnosis of leptospirosis but in Thailand microagglutination (MA) is the only serodiagnosis available. MA is not rapid and it is used mainly in the referent laboratory. In addition, its roles in early diagnosis are rarely available. Rapid screening serological test which is sensitive early in the infection is needed. Latex agglutination (LA), indirect hemagglutination (IHA) were developed and evaluated in 100 MA positive sera and 200 blood donors. Later on, IHA and LA were compared with MA in 30 patients with a clinical picture compatible with leptospirosis. IHA and LA had sensitivities of 94 and 98 per cent respectively in MA positive sera. The specificity of IHA and LA in 200 blood donors was 99 and 100 per cent respectively. The study in 30 patients showed that LA and IHA were definitely more sensitive than MA test in sera collected within two weeks after the onset of fever. LA is also one of the most rapid tests for leptospirosis. With either LA or IHA human leptospirosis will be diagnosed more readily and more accurately.