Standard Operating Procedure for the Good Manufacturing Practice-Compliant Production of Human Bone Marrow Mesenchymal Stem Cells

Roseti, Livia; Serra, Marta; Bassi, A.

doi:10.1007/7651_2014_103

Cited by 11 publications

(6 citation statements)

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“…Furthermore, some researchers advocate the use of standard operating procedures (SOPs) as being superior to the teaching of “laboratory manuals,” in that SOPs provide a step-by-step guide to the details related to a process which allows for the exact replication of all steps involved when repeating the process (Angiuoli et al, 2008; Tuck et al, 2009). It is important to note that SOPs are widely adopted in many other areas (Angiuoli et al, 2008), such as biology (Roseti et al, 2015) or medicine, for example in stroke prevention and treatment (Ntaios et al, 2015), critical illness (Sherren et al, 2014), or pre-hospital critical care interventions (Rognas et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A Review of Countermovement and Squat Jump Testing Methods in the Context of Public Health Examination in Adolescence: Reliability and Feasibility of Current Testing Procedures

Karsten²,

et al. 2019

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BackgroundIn the context of a public health physical fitness (PF) examination in adolescence, a countermovement jump (CMJ) and a squat jump (SJ) are two vertical jump (VJ) tests widely used to evaluate lower limb muscle strength and power, respectively. The main criticism of both the CMJ and SJ test is the lack of test standardization. Therefore, the objectives of this review are: (a) to gather information about both jumps; (b) to investigate whether it is possible to identify common procedures referred to in the CMJ and SJ technical execution, and (c) to design standard operating procedures (SOPs) to promote CMJ and SJ standardization in an adolescent population aged 12–18 years.MethodsThe review partially adopted the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Statement (PRISMA). Due to growing attention in monitoring physical health through field tests in recent years, articles were collected using the PubMed, Web of Science, and Scopus databases from January 2009 to July 2019. Original articles in which CMJ or SJ were used to assess the muscular strength in adolescents were eligible for further analysis. Articles written in English was imposed as a limit.ResultsA total of 117 studies met the inclusion criteria. The description of the CMJ and SJ test procedures was different within the literature, with discrepancies in the jump technique, number of jumps, and measurement devices used.ConclusionsA lack of method standardization for both the CMJ and the SJ test was identified. Based on the literature, SOPs for both VJs were proposed. These are useful in the context of public health PF examination in adolescents, as they facilitate an unbiased comparison of jump performance data between published studies.

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Section: Introductionmentioning

confidence: 99%

A Review of Countermovement and Squat Jump Testing Methods in the Context of Public Health Examination in Adolescence: Reliability and Feasibility of Current Testing Procedures

Karsten²,

et al. 2019

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“…The informed consents were obtained from the parents or guardians, and the ethics approval was required according to the guidelines from the Children’s Hospital of Zhejiang University School of Medicine Committee on the Use of Human Subjects in Research (approval number: 2013-GJ-003; approval date: February 25, 2013). BM-MSCs were prepared as described previously [10]. Briefly, 2–3 ml bone marrow specimens were mixed with an equal amount of DMEM (Invitrogen, Carlsbad, CA) supplemented with 10% of fetal bovine serum (FBS).…”

Section: Methodsmentioning

confidence: 99%

Effect of ectopic high expression of transcription factor OCT4 on the “stemness” characteristics of human bone marrow-derived mesenchymal stromal cells

et al. 2019

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Objective To investigate the effect of ectopic high expression of OCT4 on the stemness characteristics of bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods BM-MSCs were collected from three de novo acute lymphoblastic leukemia (ALL) and three aplastic anemia patients (AA), which were cultivated by the whole bone marrow adherent method. Surface markers of BM-MSCs were analyzed by flow cytometry (FCM); meanwhile, growth characteristics were observed with a phase contrast microscope, and population doubling time (PDT) was calculated. The optimal generation cells (P4) were used for the subsequent experiments. Recombinant plasmid pcDNA3.1-OCT4 was constructed and transferred into ALL MSCs by liposome transfection. The cells with stable and high expression of OCT4 were selected by G418 resistance screening and subcloning, of which the expression of OCT4 was verified by FCM, cellular immunofluorescence assay (CIFA), and RT-PCR. The expression of stemness-related transcription factors (TFs) (NANOG, SOX2) and the embryonic stem cell (ESC)-related surface markers (SSEA4, TRA-1-60, and TRA-1-81) were analyzed by FCM, RT-PCR, and CIFA. Embryonic body (EB) formation was performed with the above cells, and triembryonic differentiation marker genes were evaluated by RT-PCR. Results The primary passage of AA MSCs grew more slowly and had longer PDT (16 days on average) than ALL MSCs (10 days on average). AA MSCs presented the same typical morphology and similar expression levels of specific mesenchymal markers as ALL MSCs, whereas the latter had a much better proliferative capacity in P4 cells ( P < 0.05). Besides, the expression levels of surface markers in ALL MSCs were slightly higher than that in AA MSCs in P4, P7, and P10 cells ( P < 0.05). Cell lines with stable and high expression of OCT4 were successfully established from ALL MSCs, which were confirmed by CIFA, FCM, and RT-PCR. Compared with untransfected parental MSCs, the mean expression levels of TFs in OCT4 overexpression MSCs were increased from 0.63 ± 0.37% to 39.39 ± 1.85% (NANOG) and from 14.34 ± 2.44% to 91.45 ± 4.56% (SOX2). The average expression levels of ESC surface markers were increased from 3.33 ± 2.35%, 1.59 ± 1.29%, and 1.46 ± 0.86% to 84.98 ± 9.2%, 57.28 ± 6.72%, and 75.88 ± 7.35% respectively for SSEA-4, TRA-1-60, and TRA-1-81, which were confirmed by CIFA analysis. Moreover, the OCT4 overexpression MSCs could form EBs ex vivo and express ectoderm (TUBB3, WNT1), mesoderm (Brachyury, TBX20), and endoderm (SPARC) genes. Conclusion Ectopic high expression of transcription factor OCT4 in BM-MSCs may drive them to grow as ESC-like cells with “stemness” characteristics. Single OCT4 transfection can upregulate the expression of other stemness-related transcription factors such as NANOG and SOX2.

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“…hMSCs were prepared as described previously (13) following the standard protocol by Roseti et al (14). Bone marrow aspirates (50 ml) were obtained from the iliac crest of 8 healthy volunteer donors (20-35 years of age) at Renmin Hospital, Hubei University of Medicine (Shiyan, China), and diluted to 1:3 with Iscove's modified Dulbecco's medium (IMDM; Gibco; Thermo Fisher Scientific Inc., Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Adenovirus-mediated bone morphogenetic protein-2 promotes osteogenic differentiation in human mesenchymal stem cells in vitro

Cao

Sun

Zhang

et al. 2017

Experimental and Therapeutic Medicine

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Abstract. Delayed and failed bone union following fracture is a common clinical complication that requires treatment in orthopedics. Cell-based therapies and tissue-engineering approaches are potential therapeutic strategies for bone repair and fracture healing. However, the effect of adenovirus expressing bone morphogenetic protein-2 (Ad-BMP-2) on the osteogenic ability of human mesenchymal stem cells (hMSCs) has remained to be fully elucidated. Therefore, in the present study, hMSCs were transduced using Ad-BMP-2 to assess the effects of its application and to determine whether Ad-BMP-2 promotes the osteogenic differentiation of hMSCs. The purity of the hMSC cultures was assessed using flow cytometric analysis. In order to assess the osteogenic activity, alkaline phosphatase activity (ALP) was measured and to estimate the osteoblastic mineralization and calcification, von Kossa staining for phosphates was performed. Cells positive for Src homology 2 domain were determined to be hMSCs and the presence of CD34 was used to distinguish hematopoietic lineages. Following treatment, the Ad-BMP-2 and control group had significantly increased ALP levels (P<0.05). Compared to the blank group and the group transfected with adenoviral vector containing LacZ, the phosphate deposition in the Ad-BMP-2 group and the positive control group treated with dexamethasone was markedly increased. The results of the present study suggested that Ad-BMP-2 promotes osteogenic differentiation in hMSCs and may have a potential application in treating delayed union and nonunion following bone fracture.

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Standard Operating Procedure for the Good Manufacturing Practice-Compliant Production of Human Bone Marrow Mesenchymal Stem Cells

Cited by 11 publications

References 11 publications

A Review of Countermovement and Squat Jump Testing Methods in the Context of Public Health Examination in Adolescence: Reliability and Feasibility of Current Testing Procedures

A Review of Countermovement and Squat Jump Testing Methods in the Context of Public Health Examination in Adolescence: Reliability and Feasibility of Current Testing Procedures

Effect of ectopic high expression of transcription factor OCT4 on the “stemness” characteristics of human bone marrow-derived mesenchymal stromal cells

Adenovirus-mediated bone morphogenetic protein-2 promotes osteogenic differentiation in human mesenchymal stem cells in vitro

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