Mesenchymal stem cell isolation and characterization from human spinal ligaments

Asari, Toru; Furukawa, Ken‐Ichi; Tanaka, Sunao; Kudo, Hitoshi; Mizukami, Hiroki; Ono, Atsushi; Numasawa, Takuya; Kumagai, Gentaro; Motomura, Shigeru; Yagihashi, Soroku; Toh, Satoshi

doi:10.1016/j.bbrc.2011.12.106

Cited by 36 publications

(21 citation statements)

References 24 publications

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“…MSCs were isolated as described in the literature [10]. Briefly, collected samples were washed with phosphate-buffered saline (PBS), minced and digested with 3 mg/ml collagenase (Type5; Sigma-Aldrich, St. Louis, MO, USA) in α-modified Eagle's medium (α-MEM; Invitrogen, Carlsbad, CA, USA) at 37°C for 3 h. Nucleated cells were plated in plastic dishes (Nalge Nunc International, Rochester, NY, USA) at density of 5 × 10 5 cells/90-mm dishes.…”

Section: Cell Isolation and Culturementioning

confidence: 99%

“…The pellet was treated with chondrogenic induction medium for 21 days. Chondrogenic induction medium consisted of complete culture medium supplemented with 500 ng/ml bone morphogenetic protein (BMP) 2), 10 ng/ml transforming growth factor (TGF)-β3 (R&D Systems, Minneapolis, MN, USA) and 100 nM dexamethasone, as previously reported [10]. The pellets were evaluated in terms of diameter and pellet wet weight.…”

Section: In Vitro Differentiation and Histological Analysismentioning

confidence: 99%

“…We have focused our attention on the roles of MSCs in the process of ectopic ossification in spinal ligamentous tissues. As a first step, we isolated MSCs from human spinal ligaments and performed immunophenotypic analysis of cell surface markers [10].…”

Section: Introductionmentioning

confidence: 99%

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Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament

Harada

Furukawa

Asari

et al. 2014

Biochemical and Biophysical Research Communications

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Ectopic bone formation is thought to be responsible for ossification of the posterior longitudinal ligament of the spine (OPLL). Mesenchymal stem cells (MSCs) were isolated from spinal ligaments and shown to play a key role in the process of ectopic ossification. The purpose of this study was to explore the capacity of these MSCs to undergo lineage commitment and to assess the gene expression changes between these committed and uncommitted MSCs between OPLL and non-OPLL patients. Spinal ligament-derived cells were obtained from OPLL patients or patients with cervical spondylotic myelopathy (non-ossified) for comparison (n=8 in each group). MSCs from the two patient cohorts were evaluated for changes in colony forming ability; osteogenic, adipogenic and chondrogenic differentiation potential; and changes in gene expression following induction with lineage-specific conditions. We show that the osteogenic differentiation potential was significantly higher in MSCs from OPLL patients than in those from non-OPLL patients. In addition, alkaline phosphatase activity and several osteogenic-related genes expressions (bone morphogenetic protein 2, runt-related transcription factor 2 and alkaline phosphatase) were significantly higher in the OPLL group than in the non-OPLL group. However, single cell cloning efficiency, adipogenic and chondrogenic differentiation, and the expression of adipogenic and 2 chondrogenic-related genes were equivalent between MSCs harvested from OPLL and non-OPLL patient samples. These findings suggest an increase in the osteogenic differentiation potential of MSCs from OPLL patients and that this propensity toward the osteogenic lineage may be a causal factor in the ossification in these ligaments.

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Section: Cell Isolation and Culturementioning

confidence: 99%

Section: In Vitro Differentiation and Histological Analysismentioning

confidence: 99%

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Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament

Harada

Furukawa

Asari

et al. 2014

Biochemical and Biophysical Research Communications

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“…It has been reported that MSC could be isolated from a variety of tissues of the body [21]. However, whether MSC are located in ACL remnants in situ is not clear yet.…”

Section: Discussionmentioning

confidence: 99%

Mesenchymal stem cells reside in anterior cruciate ligament remnants in situ

Tang

et al. 2015

International Orthopaedics (SICOT)

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“…They have thus gained attention as a cellular source for regenerative medicine applications (43). However, it has been suggested that these cells may work in undesirable ways in pathological conditions, such as in aortic valve calcification (44,45), fibrodysplasia ossificans progressiva (FOP) (46), ectopic ossification in spinal ligaments (47,48), and ectopic calcification following burn injury (49). Mesenchymal stem cells can cause improper calcification and ossification after differentiating into chondrocyte-or osteoblast-like cells according to environmental and local factors (48 -50 (28).…”

Section: What Are the Cellular Origins In Calcification?mentioning

confidence: 99%

Recent Advances in Research on Human Aortic Valve Calcification

Furukawa

2014

J Pharmacol Sci

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Abstract. Aortic valve calcification can aggravate aortic stenoses, and it is a significant cause of sudden cardiac death. The increasing number of patients with age-related calcification is a problem in developed nations. However, the only treatment option currently available is highly invasive cardiac valve replacement. Therefore, clarification of the etiology of calcification is urgently needed to develop drug therapies and prevention methods. Recent studies have revealed that calcification is not a simple sedimentation of a mineral through a physicochemical phenomenon; various factors dynamically contribute to the mechanism. Further, we are finally beginning to understand the cellular origins of calcification, which had been unclear for a long time. Based on these findings that help to clarify potential drug targets, we expect to establish drug therapies that reduce the stress on patients. In this paper, I introduce the latest findings on cells that are most likely to contribute to calcification and on calcification-related factors that may lead to the development of drug therapies.

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Mesenchymal stem cell isolation and characterization from human spinal ligaments

Cited by 36 publications

References 24 publications

Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament

Osteogenic lineage commitment of mesenchymal stem cells from patients with ossification of the posterior longitudinal ligament

Mesenchymal stem cells reside in anterior cruciate ligament remnants in situ

Recent Advances in Research on Human Aortic Valve Calcification

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