• Title/Summary/Keyword: tissue cultures

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Clinical Application of Adipose Derived Stromal Cell Autograft for Wound Coverage (배양하지 않은 지방조직세포를 이용한 창상피복)

  • Seo, Dong-lin;Han, Seung-Kyu;Chun, Kyung-Wook;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.35 no.6
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    • pp.653-658
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    • 2008
  • Purpose: Skin and soft tissue defect is one of the major challenges faced by plastic surgeons. Adipose derived stromal cells, which can be harvested in large quantities with low morbidity, display multilineage mesodermal potential. Therefore, adipose derived stromal cells have been met with a great deal of excitement by the field of tissue engineering. Recently, Adipose derived stromal cells have been isolated and cultured to use soft tissue restoration. In order to apply cultured cells for clinical purpose, however, FDA approved facilities and techniques are required, which may be difficult for a clinician who cultures cells in a laboratory dedicated to research to utilize this treatment for patients. In addition, long culture period is needed. Fortunately, adipose derived stromal cells are easy to obtain in large quantities without cell culture. The purpose of this study is to present a possibility of using uncultured adipose derived stromal cells for wound coverage. Methods: Seven patients who needed skin and soft tissue restoration were included. Five patients had diabetic foot ulcers, 1 patient got thumb amputation, and 1 patient had tissue defect caused by resection of squamous cell carcinoma. The patients' abdominal adipose tissues were obtained by liposuction. The samples were digested with type I collagenase and centrifuged to obtain adipose derived stromal cells. The isolated adipose derived stromal cells were applied over the wounds immediately after the wound debridement. Fibrin was used as adipose derived stromal cells carrier. Occlusive dressing was applied with films and foams and the wounds were kept moist until complete healing. Results: One hundred to one hundred sixty thousand adipose derived stromal cells were isolated per ml aspirated adipose tissue. All patients' wounds were successfully covered with the grafted adipose derived stromal cells in a 17 to 27 day period. No adverse events related to this treatment occurred. Conclusion: The use of uncultured adipose derived stromal cells was found to be safe and effective treatment for wound coverage without donor site morbidity.

Nontuberculous Mycobacterial Infection after Body Contouring Procedure, Case Report (체형 교정 시술 후 발생한 비정형 마이코박테리아 감염, 증례 보고)

  • Jeong, Jae-Yeon;Lim, So-Young;Pyon, Jai-Kyong;Mun, Goo-Hyun;Bang, Sa-Ik;Oh, Kap-Sung
    • Archives of Plastic Surgery
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    • v.37 no.3
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    • pp.293-296
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    • 2010
  • Purpose: NTM (non tuberculous mycobacteria) is rare cause of surgical site infection after plastic surgery in immunocompetent patients. There are some reports about NTM infection after body contouring procedure from Latin America. But, there is no report in Korea. The purpose of this article is to report 2 patients with soft tissue infection caused by NTM after body contouring procedure. Methods: Two young female patients exhibited signs of inflammation and abscess after body contouring procedure. One patient underwent liposuction. The other underwent HPL (hypotonic pharmacologic lipo-dissolution) injection. Results: The result of tissue cultures were positive for NTM. All patients responded to the combined therapeutic approach. Conclusion: The goal of this article is to raise awareness among plastic surgeons who may encounter such patients in their practice. NTM should be included in the differential diagnosis of surgical site infection after body contouring surgery.

Studies on tissue culture of medicinal plants (II) -Tissue cultures of Glycyrrhiza glabra L. var. glandulifera $R_{EG.}$ et $H_{ERDER}$- (약용식물(藥用植物)의 조직배양(組織培養)에 관(關)한 연구(硏究)(II) -이태리감초(甘草)의 조직배양(組織培養)-)

  • Yoo, Sung-Cho;Kim, Sung-Soon
    • Korean Journal of Pharmacognosy
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    • v.7 no.1
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    • pp.55-57
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    • 1976
  • The callus formation of Glycyrrhiza glabra L. var. $glandulifera\;R_{EG}.\;et\;H_{ERD}$. in tissue culture was promoted on Murashige and Skoog's basal solution supplemented with 40g/l of sucrose, 1mg/l of kinetin and 5mg/l of 2, 4-D. The fresh and dry weights of callus and glycyrrhizin contents in callus of the Glycyrrhiza glabra L. var. $glandulifera\;R_{EG}.\;et\;H_{ERD}$. were determined monthly up to 12 months and obtained the results as follows: 1.The fresh weight of formed callus was increased rapidly from 2 to 4 months but growing rate of callus was slow from 4 to 6 months. This indicates that the cell division of callus was most active during the first $2{\sim}3$ months. 2. Glycyrrhizin contents in callus were also increased but the contents were not related to the increased weight of callus.

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Immunohistochemistry for detection of Aujeszky's disease virus antigens : Immunogold-silver method in tissue sections (오제스키병 바이러스 항원검출을 위한 면역조직화학적 연구 -조직절편내 immunogold-silver법-)

  • Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.365-369
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    • 1988
  • The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

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Anatomical Observation of Vitrified and Glaucous Leaf from Rehmannia glutinosa Plant Produced in Vitro (지황 기내배양시 투명화된 잎과 정상잎간의 조직학적 관찰)

  • 백기엽;유광진;박상일;신성련
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.323-327
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    • 1997
  • Addition of growth inhibitors such as ancymidol, ABA, chloromequat, and pachlobutrazol into MS medium had no effect to preventing vitrification in cultures of Rehmania glutinosa. Anatomical investigation revealed that vitrified thick leaf tissue in vitro had larger intercellular space with poor development of sponge and pallisade tissue compared to those of in vitro grown glaucous and field grown plants. In vitro grown glaucous leaf had smaller and round type stomata showing distinguishable guard and subsidiary cell than those of reestablished plantlets into soil whereas abnormal stomata and poor development of epicuticular wax on the surface of leaf was observed in verified plantlet.

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High Dose of FGF-2 Induced Growth Retardation via ERK1/2 De-phosphorylation in Bone Marrow-derived Mesenchymal Stem Cells

  • Shim, Kwang Yong;Saima, Fatema Tuj;Eom, Young Woo
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.49-56
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    • 2017
  • Fibroblast growth factor (FGF)-2 is one of the most effective growth factors to increase the growth rate of mesenchymal stem cells (MSCs). Previously, we reported that low dose of FGF-2 (1 ng/ml) induced proliferation of bone marrow-derived mesenchymal stem cells (BMSCs) through AKT and ERK activation resulting in reduction of autophagy and senescence, but not at a high dose. In this study, we investigated the effects of high dose FGF-2 (10 ng/ml) on proliferation, autophagy and senescence of BMSCs for long term cultures (i.e., 2 months). FGF-2 increased the growth rate of BMSCs in a dose dependent manner for a short term (3 days), while during long term cultures (2 months), population doubling time was increased and accumulated cell number was lower than control in BMSCs when cultured with 10 ng/ml of FGF-2. 10 ng/ml of FGF-2 induced immediate de-phosphorylation of ERK1/2, expression of LC3-II, and increase of senescence associated ${\beta}$-galactosidase (SA-${\beta}$-Gal, senescence marker) expression. In conclusion, we showed that 10 ng/ml of FGF-2 was inadequate for ex vivo expansion of BMSCs because 10 ng/ml of FGF-2 induced growth retardation via ERK1/2 de-phosphorylation and induction of autophagy and senescence in BMSCs.

Effect of Cytokines and bFGF on the Osteoclast Differentiation Induced by $1\;{\alpha},25-(OH)_2D_3$ in Primary Murine Bone Marrow Cultures

  • Chae, Han-Jung;Kang, Jang-Sook;Bang, Byung-Gwan;Cho, Seoung-Bum;Han, Jo-Il;Choi, Joo-Young;Kim, Hyung-Min;Chae, Soo-Wan;Kim, Hyung-Ryong
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.539-546
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    • 1999
  • Bone is a complex tissue in which resorption and formation continue throughout life. The bone tissue contains various types of cells, of which the bone forming osteoblasts and bone resorbing osteoclasts are mainly responsible for bone remodeling. Periodontal disease represents example of abnormal bone remodeling. Osteoclasts are multinucleated cells present only in bone. It is believed that osteoclast progenitors are hematopoietic origin, and they are recruited from hematopoietic tissues such as bone marrow and circulating blood to bone. Cells present in the osteoclast microenvironment include marrow stromal cells, osteoblasts, macrophages, T-lymphocytes, and marrow cells. These cells produce cytokines that can affect osteoclast formation. In vitro model systems using bone marrow cultures have demonstrated that $IL-l{\beta},\;IL-3,\;TNF-{\alpha},$ bFGF can stimulate the formation of osteoclasts. In contrast, IL-4 inhibits osteoclast formation. Knowledge of cytokines and bFGF that affect osteoclast formation and their capacity to modulate the bone-resorbing process should provide critical insights into normal calcium homeostasis and disorders of bone turnover such as periodontal disease, osteoporosis and Paget's disease.

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Rhamnose-rich and fucose-rich oligo- and polysaccharides (RROP-s and FROPs), agonists and antagonists of cell-membrane receptors as new active principles against skin aging.

  • Robert, L.;Robert, A.M.;Gesztes, J.L.;Luppi, E.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.352-373
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    • 2003
  • Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.

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Effect of Plant Growth Regulators on Multiple Shoot Formation and Elongation from Shoot Tip Cultures of Grape Species (포도의 경정배양에 의한 다아체형성 및 신장에 미치는 생장조절제의 영향)

  • 서정해;정재동;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.25-32
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    • 2001
  • Shoot tips of grape were cultured in uitro and tried to identify optimal culture conditions for regeneration, multiple shoot formation from meristemoid tissue and those subsequent elongation of multi-shoots. Healthy growing shoots were taken in early May, rinsed with running tap water, soaked in a neutral detergent and washed with soft brushing, and washed out with tap water, then sterilized with 10g Ca(ClO)$_2$/140 mL distilled water (Wilson's solution) for 5 min. Survival percentage of the cultures which were sterilized as above procedures was highly increased, compared with the other sterilized method. Propagation of multi-shoots from meristemoid showed a good response in 3/4 strength MS medium enriched with 0.1 mg/L NAA and 3.0 mg/L BA. Shoot elongation from multi-shooting clump well occurred in 3/4 strength MS medium supplemented with 80 mg/L adenine sulfate, 0.1 mg/L NAA and 1.0~2.0 mg/L BA.

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Production of ${\gamma}$-Linolenic Acid by Cell Suspension Cultures of Lithospermum erythrorhizon (지치세포 배양에 의한 ${\gamma}$-Linolenic Acid 생산)

  • 김용환;김정봉;류태훈;이철희;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.111-114
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    • 1995
  • To produce ${\gamma}$-linolenic acid (GLA) by cell cultures of Lithospermum erythrorhizon, we optimized medium compositions including carbon sources, nitrogen sources and growth regulators. MS basal medium supplemented with 1.0 mg/L 2, 4-D was effective for callus induction from mesophyll tissue. Addition of sucrose at 88mM concentration induced active proliferation of suspension cells and increased GLA content. Increased supplement of potassium nitrate as nitrogen source resulted in proliferous cell growth and increased total fatty acid content Abscisic acid increased cell growth and fatty acid content in callus culture, whereas as it had an inhibitory effect in suspension cell culture.

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