• Nutritional Sciences
• /
• v.7 no.3
• /
• pp.144-150
• /
• 2004

We have investigated the effect of calcium spirulan(Ca-SP) isolated from a blue-green alga Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose and fructose, on invasion of both B16- BL6 melanoma cells, Colon 26 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters in a concentration-dependent manner. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no inhibitory effect on tumor cell migration to fibronectin-coated filters. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel- and laminin-substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin in a concentration-dependent fashion, while the pretreatment of laminin-substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contraset, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP in a dose-dependent manner. Seven intermittent ⅰ.ⅴ. injection of 100$\mu\textrm{g}$ of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung colonization of B16-BL6 melanoma cells in experimental metastasis model, by inhibiting the tumor invasion of basement membrane Matrigel, probably through the prevention of the adhesion and migration of tumor cells to laminin-substrate and of the heparanase activity.

• Journal of Veterinary Clinics
• /
• v.29 no.5
• /
• pp.412-415
• /
• 2012

A 1-year-old, castrated male Yorkshire terrier (case 1) and a 7-year-old female Poodle (case 2) presented with delayed union fractures. In case 1, the dog had a fractured right distal radius and ulna. In case 2, the dog had a fractured left distal tibia and fibula. A physical examination and radiographs performed in both dogs revealed delayed union fractures with large gaps. The fracture sites were fixed by bone plate and screws. Autogenous cancellous bone graft was applied into the fracture gap. To encourage rapid bone union, we used matrigel containing $20{\mu}g$ of recombinant human bone morphogenetic protein-2 (rhBMP-2) in the fracture site. Radiographs were taken postoperatively to monitor healing. Rapid bone union was noted in both dogs in long-term radiographs. In case 1, the radiographs revealed that the fracture gaps of the radius and ulna were bridged at 2 weeks. Fracture lines were not observed and normal appearance was restored at 20 weeks. In case 2, the radiographs showed that fracture repair had progressed at 11 weeks. The fractures healed faster than expected in these two cases. The results indicate that rhBMP-2 and matrigel may be effective and useful materials to enhance healing of delayed fractures.

• Proceedings of the Korean Society of Developmental Biology Conference
• /
• 2001.08a
• /
• pp.38-38
• /
• 2001

TM4 Sertoli cell의 체외 관강형성 유도에 미치는 세포외기질 (ECM) 및 hepatocyte growth factor (HGF)의 역할과 세포분화 과정에서 MMP의 발현의 변화를 조사하였다. Matrigel bed(60%, v/v) 상에서 배양한 TM4 cell은 무혈청 조건하에서 chain 분화단계를 거쳐 cord의 구조로 분화하였다. 그러나 이후의 분화는 일어나지 않았다. TM4 cell에서 c-MET (HGF receptor)의 발현을 확인하였으며 HGF를 첨가한 배양액에서 분화가 촉진되었으며, cord에서 tubule로의 분화가 유도되었다. 또한 TM4 cell의 분화는 MMP-2 및 MMP-9의 발현이 증가를 수반하였으며 HGF는 MMPs의 발현을 증가시켰다. GFR-Matrigel과 성장인자인 HGF는 무혈청 배지에서 TM4 cell의 체외에서 관강형성에 필요한 환경을 제공하며, MMP-2 및 -9은 TM4 cell의 체외분화 과정에서 조절역할을 수행하는 것으로 사료된다.

• Molecules and Cells
• /
• v.37 no.6
• /
• pp.473-479
• /
• 2014

Spermatogonial stem cells (SSCs, also called germline stem cells) are self-renewing unipotent stem cells that produce differentiating germ cells in the testis. SSCs can be isolated from the testis and cultured in vitro for long-term periods in the presence of feeder cells (often mouse embryonic fibroblasts). However, the maintenance of SSC feeder culture systems is tedious because preparation of feeder cells is needed at each subculture. In this study, we developed a Matrigel-based feeder-free culture system for long-term propagation of SSCs. Although several in vitro SSC culture systems without feeder cells have been previously described, our Matrigel-based feeder-free culture system is time- and cost-effective, and preserves self-renewability of SSCs. In addition, the growth rate of SSCs cultured using our newly developed system is equivalent to that in feeder cultures. We confirmed that the feeder-free cultured SSCs expressed germ cell markers both at the mRNA and protein levels. Furthermore, the functionality of feeder-free cultured SSCs was confirmed by their transplantation into germ cell-depleted mice. These results suggest that our newly developed feeder-free culture system provides a simple approach to maintaining SSCs in vitro and studying the basic biology of SSCs, including determination of their fate.

• Journal of Acupuncture Research
• /
• v.22 no.2
• /
• pp.171-180
• /
• 2005

Background & Objective : Angiogenesis consists of the proliferation, migration, and differentiation of endothelial cells, and angiogenic factors and matrix protein interactions modulate this process. The aim of this study was to determine whether Puerariae radix could induce angiogenic activity in human umbilical vein endothelial cells (HUVECs). Methods: The angiogenic activity of Puerariae radix were evaluated by using BrdU assay, chemotactic migration assay, tube formation assay, measurement of bFGF in HUVECs, and Matrigel plug assay in mice. Results : Puerariae radix significantly increased HUVECs proliferation in a dose-dependent manner. In addition, Puerariae radix increased migration and tube-like formation in HUVECs. Interestingly,the expression of basic fibroblast growth factor (bFGF), an angiogenesis-stimulating growth factor, was dose-dependently increased by Puerariae radix. The angiogenic activity of Puerariae radix was confirmed using an in vivo Matrigel angiogenesis model, showing promotion of blood vessel formation. Conclusion : Puerariae radix significantly induces angiogenesis in vitro and in vivo. These results suggest that Puerariae radix is a potent angiogenic agent, and a promising drug, for the induction of neovascularization.

• Archives of Pharmacal Research
• /
• v.21 no.4
• /
• pp.398-405
• /
• 1998

We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. $G_2$/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of $G_1$ phase cells was not observed. The colonies with distinct multicelfular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.30 no.1
• /
• pp.1-16
• /
• 2004

The use of artificial nerve conduit containing viable Schwann cells is one of the most promising strategies to repair the peripheral nerve injury. To fabricate an effective nerve conduit whose microstructure and internal environment are more favorable in the nerve regeneration than existing ones, a new three-dimensional Schwann cell culture technique using $Matrigel^{(R)}$. and dorsal root ganglion (DRG) was developed. Nerve conduit of three-dimensionally arranged Schwann cells was fabricated using direct seeding of freshly harvested DRG into a $Matrigel^{(R)}$ filled silicone tube (I.D. 1.98 mm, 14 mm length) and in vitro rafting culture for 2 weeks. The nerve regeneration efficacy of three-dimensionally cultured Schwann cell conduit (3D conduit group, n=6) was assessed using SD rat sciatic nerve defect of 10 mm, and compared with that of silicone conduit filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method (2D conduit group, n=6). After 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were examined using image analyzer and electromicroscopic methods. The SFI and ankle stance angle (ASA) in the functional evaluation were $-60.1{\pm}13.9$, $37.9^{\circ}{\pm}5.4^{\circ}$ in 3D conduit group (n=5) and $-87.0{\pm}12.9$, $32.2^{\circ}{\pm}4.8^{\circ}$ in 2D conduit group (n=4), respectively. And the myelinated axon was $44.91%{\pm}0.13%$ in 3D conduit group and $13.05%{\pm}1.95%$ in 2D conduit group to the sham group. In the TEM study, 3D conduit group showed more abundant myelinated nerve fibers with well organized and thickened extracellular collagen than 2D conduit group, and gastrocnemius muscle and biceps femoris tendon in 3D conduit group were less atrophied and showed decreased fibrosis with less fatty infiltration than 2D conduit group. In conclusion, new three-dimensional Schwann cell culture technique was established, and nerve conduit fabricated using this technique showed much improved nerve regeneration capacity than the silicone tube filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method.

• Proceedings of the KSMRM Conference
• /
• 2007.10a
• /
• pp.46-46
• /
• 2007

• Biomaterials and Biomechanics in Bioengineering
• /
• v.2 no.1
• /
• pp.33-45
• /
• 2015

To engineer tissue-like structures, cells are required to organize themselves into three-dimensional networks that mimic the native tissue micro-architecture. Here, we present agarose-based multi-well platform incorporated with electrical stimulation to build skeletal muscle-like tissues in a facile and highly reproducible fashion. Electrical stimulation of C2C12 cells encapsulated in collagen/matrigel hydrogels facilitated the formation 3D muscle tissues. Consequently, we confirmed the transcriptional upregulations of myogenic related genes in the electrical stimulation group compared to non-stimulated control group in our multi-well 3D culture platform. Given the robust fabrication, engineered muscle tissues in multi-well platform may find their use in high-throughput biological studies drug screenings.

• Korean Journal of Oriental Medicine
• /
• v.4 no.1 s.4
• /
• pp.129-138
• /
• 1998

Angiogenesis, the formation of new blood vessels, is considered to be involved in many pathological symptoms such as diabetic retinopathy, arthritis, inflammation and solid tumour. In particular, it is thought that angiogenesis is critical for development and growth of solid tumour. Recent study shows that there is a highly significant association of microvessel density with overall survival and relapse-free survival in patients with breast tumour In this study, the inhibition effect of angiogenesis of traditional herbs used for the treatment of cancer was examined. It was found out that the extract of Agaricus blazei by boiling water is a possible inhibitor of angiogenesis. It inhibited normal developmental angiogenesis In the chorioallantoic membrane of chick embryos and also inhibited capillary-like tube formation by endotherial cells on matrigel in vitro. These results suggest that Agaricus blazei can be a potent angiogenesis inhibitor.