Isolation and Characterization of Mouse Mesenchymal Stem Cells

Sung, Jong Hwan; Yang, Hyung-In; Park, J.B.; Choi, Gwang Seong; Joh, Jae‐Won; Kwon, Choon Hyuck David; Chun, Jiang; Lee, S.-K.; Kim, S.-J.

doi:10.1016/j.transproceed.2008.08.009

Cited by 141 publications

(111 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Observa-se crescimento rápido inicial até 72 horas, fase exponencial entre 72 e 192 horas, seguida de platô por saturação da densidade. Estudos com CTMs de humanos descreveram as células em cultivo como células de morfologia fibroblástica (Barry e Murphy, 2004;Dvorakova et al, 2008), assim como estudos realizados com CTMs de camundongos (Sung et al, 2008;Tropel et al, 2004) e de cães (Csaki et al, 2007). Porém, em nenhum dos estudos com avaliação da morfologia das CTMs foi realizado o estudo morfométrico.…”

Section: Resultsunclassified

Isolamento e caracterização de células mesenquimais do tecido adiposo de cães

Patricio

Rebelatto

Brofman

et al. 2013

Arq. Bras. Med. Vet. Zootec.

View full text Add to dashboard Cite

RESUMOAs células-tronco mesenquimais (CTMs) diferenciam-se em várias linhagens e têm potencial de utilização na medicina regenerativa. As CTMs podem ser isoladas de vários tecidos de animais adultos. O objetivo deste estudo foi o isolamento das CTMs do tecido adiposo de cães, seu cultivo e diferenciação. Foram coletadas amostras de tecido adiposo subcutâneo de cinco cães. As CTMs foram isoladas, obtendo-se 146.803 (±49.533) células/g, cultivadas e diferenciadas em osteoblastos, adipócitos e condrócitos. Avaliaram-se a cinética do crescimento, a morfologia e a viabilidade celular. A caracterização citoquímica comprovou a natureza mesenquimal das células isoladas. O cultivo foi iniciado com 20.000 células/mL, verificando-se crescimento rápido até 72 horas (220.000 células/mL), fase exponencial entre 72 e 192 horas (455.000 células/mL), seguida de platô por saturação da densidade com 240 horas (355.000 células/mL). A viabilidade celular variou entre 96 e 100%. As CTMs em cultivo são fibroblásticas, fusiformes, com citoplasma basofílico e núcleo esférico. O comprimento médio das células variou entre 80,85 e 98,36µm, a largura média entre 17,40 e 28,79µm e o diâmetro médio do núcleo entre 15,46 e 17,74µm.Palavras-chave: morfologia celular, diferenciação celular, células-tronco, panículo adiposo 80.85-98.36µm, 17.40-28.79µm and 15.46-17.74µm respectively. ABSTRACT The applications of mesenchymal stem cells (MSCs

show abstract

Section: Resultsunclassified

Isolamento e caracterização de células mesenquimais do tecido adiposo de cães

Patricio

Rebelatto

Brofman

et al. 2013

Arq. Bras. Med. Vet. Zootec.

View full text Add to dashboard Cite

show abstract

“…25,89,160 Donor-related differences also contribute to the variance, 161 as do species-related, and even strain-related, differences. 89,162 Differences among MSC isolated from different tissues 161,163 may reflect the diversity among MSC niches. 164 Exposure of MSCs to a chemoattractant may stimulate collateral responses in addition to the chemotaxis desired.…”

Section: Discussionmentioning

confidence: 99%

In SituTissue Regeneration: Chemoattractants for Endogenous Stem Cell Recruitment

Berg-Foels

2014

Tissue Engineering Part B: Reviews

131

111

View full text Add to dashboard Cite

Tissue engineering uses cells, signaling molecules, and/or biomaterials to regenerate injured or diseased tissues. Ex vivo expanded mesenchymal stem cells (MSC) have long been a cornerstone of regeneration therapies; however, drawbacks that include altered signaling responses and reduced homing capacity have prompted investigation of regeneration based on endogenous MSC recruitment. Recent successful proof-of-concept studies have further motivated endogenous MSC recruitment-based approaches. Stem cell migration is required for morphogenesis and organogenesis during development and for tissue maintenance and injury repair in adults. A biomimetic approach to in situ tissue regeneration by endogenous MSC requires the orchestration of three main stages: MSC recruitment, MSC differentiation, and neotissue maturation. The first stage must result in recruitment of a sufficient number of MSC, capable of effecting regeneration, to the injured or diseased tissue. One of the challenges for engineering endogenous MSC recruitment is the selection of effective chemoattractant(s). The objective of this review is to synthesize and evaluate evidence of recruitment efficacy by reported chemoattractants, including growth factors, chemokines, and other more recently appreciated MSC chemoattractants. The influence of MSC tissue sources, cell culture methods, and the in vitro and in vivo environments is discussed. This growing body of knowledge will serve as a basis for the rational design of regenerative therapies based on endogenous MSC recruitment. Successful endogenous MSC recruitment is the first step of successful tissue regeneration

show abstract

Section: Acceleration Of Adipocyte Differentiation In Mesenchymal Stementioning

confidence: 80%

“…We confirmed that GATA2 fl BM-MSC retained the potential to differentiate into adipogenic lineages (Online Supplementary Figure S1B). Flow cytometric analysis confirmed the characteristic immunophenotype, 28 showing that GATA2 fl BM-MSC expressed CD29, CD44 and Sca-1 but not markers such as CD11b, CD34 and CD45 (Online Supplementary Figure S1C).To determine whether the loss of GATA2 influenced the BM-MSC phenotype, the DNA-binding domain of GATA2 was deleted using the Cre-loxP system, and GATA2 knockout-BM-MSC (GATA2 − BM-MSC) were generated. Quantitative RT-PCR analysis revealed that Gata2 expression was significantly decreased in the GATA2 -MSC, implying that iCre-mediated deletion of the GATA2 C-finger resulted in decreased GATA2 autoregulation ( Figure 1A).…”

mentioning

confidence: 92%

GATA2 regulates differentiation of bone marrow-derived mesenchymal stem cells

et al. 2014

View full text Add to dashboard Cite

Methods Generation of bone marrow mesenchymal stem cellsTo generate mouse BM-MSC, bone marrow cells from GATA2 conditional knockout mice were cultured in MesenCult MSC Basal Medium supplemented with 20% MSC stimulatory supplements (Stem Cell Technologies). The BM-MSC were transfected with the retroviruses expressing iCre to delete the DNA binding domain of GATA2 by inducing the Cre-loxP system. 21,22 To generate human BM-MSC, bone marrow mononuclear cells from healthy donors were cultured with Dulbecco's modified Eagle's medium (Life Technologies) supplemented with 20% fetal bovine serum (Life Technologies), 10 ng/mL basic fibroblast growth factor (PeproTech), 10 mM HEPES (Life Technologies), and 100 μg/mL penicillin/streptomycin (Invitrogen). [23][24][25] Established BM-MSC were used until the seventh generation.The study was approved by the ethical committee of Tohoku University Graduate School of Medicine. Clinical samples were collected after obtaining written informed consent. The ethics policies of the Declaration of Helsinki were followed. Characterization of bone marrow mesenchymal stem cellsBM-MSC immunophenotypes were determined using a FACSAria II (BD). To induce differentiation into adipocytes, human Mesenchymal Stem Cell Adipogenic Differentiation Medium (Lonza) was used. After 12-16 days, morphological changes were assessed using an inverted microscope. Typical adipocytes were stained with Oil Red O.2 The area of mature adipocytes was determined using HistoQuest software (Novel Science). Quantitative reverse transcriptase polymerase chain reaction analysis and transcription profilingQuantitative reverse transcriptase polymerase chain reaction analysis (RT-PCR) was performed as previously described.26 Primer sequences are available upon request.For transcription profiling, the Human Genome U133 Plus 2.0 Array was used (Affymetrix). Gene ontology analysis was conducted using the DAVID bioinformatics program (http://david.abcc.ncifcrf.gov/). Short interfering RNA-mediated knockdownAnti-GATA2 and control short interfering RNA (siRNA) 26 were transfected into human BM-MSC with Lipofectamine TM RNAiMAX reagent (Life Technologies). Cells were analyzed 48 h after transfection. Viral vectors and cell transductionRetroviral overexpression of GATA2 was performed using the MSCV retrovirus vector, which co-expresses green fluorescent protein (GFP) by internal ribosome entry sites (IRES), transfecting into Platinum Retroviral Packaging Cell Lines (PLAT-F) 27 with FuGENE HD (Roche). Human BM-MSC were pretreated with Retronectin (TAKARA BIO.), and GFP-positive cells were sorted using FACSAria II (BD Biosciences).Co-culture of CD34-positive-enriched cells with a mesenchymal stem cell feeder layer BM-MSC were transfected with control or GATA2-siRNA. On day 3, control and GATA2 knockdowned BM-MSC, respectively, were replaced with serum-free medium containing CD34-positiveenriched cells (RIKEN). Serum-free medium (StemPro-34 SFM: Life Technologies) contained 100 ng/mL stem cell factor, 100 ng/mL interleukin (...

show abstract

Isolation and Characterization of Mouse Mesenchymal Stem Cells

Cited by 141 publications

References 19 publications

Isolamento e caracterização de células mesenquimais do tecido adiposo de cães

Isolamento e caracterização de células mesenquimais do tecido adiposo de cães

In SituTissue Regeneration: Chemoattractants for Endogenous Stem Cell Recruitment

GATA2 regulates differentiation of bone marrow-derived mesenchymal stem cells

Contact Info

Product

Resources

About