九叔归来3魁蛊婴在线观看_男人躁女人到高潮AV_香港成人论坛_亚洲精品久久久久久偷窥_夜来香成人网_亚洲制服 视频在线观看_无毒黄站_国产传媒18精品A片一区_麻花豆传媒剧国产MV在线观看_东北60岁熟女露脸在线_国产高清视频在线观看97_一道本视频一二三区_yellow免费播放在线观看_浪漫樱花动漫在线观看官网_高清AV熟女一区_天堂在线www_亚洲第一成年人网站_黄色在线免费观看_av女优快播_久久精品99国产精品日本

English | 中文版 | 手機版 企業(yè)登錄 | 個人登錄 | 郵件訂閱
當前位置 > 首頁 > 技術文章 > Isolation and cultivation of endothelial progenitor cells (EPCs)

Isolation and cultivation of endothelial progenitor cells (EPCs)

瀏覽次數(shù):3401 發(fā)布日期:2011-9-30  來源:www.pricells.com.cn
           Isolation and cultivation of endothelial progenitor cells (EPCs)      

Circulating bone marrow (BM)–derived endothelial progenitor cells (EPCs) are recruited to the site of tissue regeneration and substantially contribute to neovascularization and re-endothelialization after acute vascular injury. Intravenous infusion of EPCs after ischemia was shown to improve neovascularization and cardiac function in both animal models and pilot clinical studies. Conversely, BM-derived EPCs participate in the pathogenesis of various diseases, such as cancer, retinopathy, and atherosclerosis.  Impaired recruitment of BM-derived endothelial and hematopoietic precursor cells is known to block both tumor angiogenesis and growth.  Injection of BM cells promotes injury-associated retinal angiogenesis. BM-derived progenitors also contribute to the formation of the microvasculature in allograft arteriosclerotic lesions.  Therefore, EPCs have both physiologic and pathologic roles; thus, they have been widely considered for establishing new therapeutic strategies for the treatment of angiogenesis-related diseases.  The number of circulating EPCs may limit the ultimate magnitude of angiogenesis therapies and, therefore, strategies that are based on the administration of ex vivo–expanded populations of EPCs harvested from the patient's circulating blood appear promising.
1. Human umbilical cord blood (HUCB) samples (∼50 mL each) were collected from fresh placentas with attached umbilical cords by gravity flow.

2. EPCs were isolated by density gradient centrifugation over Biocoll (Biochrom, Berlin, Germany) for 30 minutes at 400g and washed 3 times in PBS (Biochrom).

3. One million cells were seeded onto 6-well plates coated with human fibronectin in Primary endothelial basal medium.

4. The medium was supplemented with endothelial growth medium containing fetal bovine serum, human VEGF-A, human fibroblast growth factor-B, human epidermal growth factor, insulin-like growth factor 1 (IGF1), and ascorbic acid in appropriate amounts.

5. After 3 days, non-adherent cells were removed, and fresh culture medium was applied.

6. Cultures were maintained for 7 days.

7. Phenotypic analyses of the cells were performed on days 3, 5, and 7.

8. EPC identification and estimation of culture purity (90%-95%) were determined by staining cells with FITC-labeled Ulex europaeus lectin I and uptake of Dil-conjugated acetylated low-density lipoprotein.

References

1. Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997;275:964-967.
2. Sata M. Role of circulating vascular progenitors in angiogenesis, vascular healing, and pulmonary hypertension: lessons from animal models. Arterioscler Thromb Vasc Biol 2006;26:1008-1014.
3. Orimo A, Gupta PB, Sgroi DC, et al. Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 2005;121:335-348.
4. Lyden D, Hattori K, Dias S, et al. Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med 2001;7:1194-1201.
5. Otani A, Kinder K, Ewalt K, Otero FJ, Schimmel P, Friedlander M. Bone marrow-derived stem cells target retinal astrocytes and can promote or inhibit retinal angiogenesis. Nat Med 2002;8:1004-1010.
6. Hu Y, Davison F, Zhang Z, Xu Q. Endothelial replacement and angiogenesis in arteriosclerotic lesions of allografts are contributed by circulating progenitor cells. Circulation 2003;108:3122-3127.
7. Assmus B, Schachinger V, Teupe C, et al. Transplantation of Progenitor Cells and Regeneration Enhancement in Acute Myocardial Infarction (TOPCARE-AMI). Circulation 2002;106:3009-3017.

發(fā)布者:武漢原生原代生物醫(yī)藥科技有限公司
聯(lián)系電話:027-87490190
E-mail:service@pricells.com.cn

用戶名: 密碼: 匿名 快速注冊 忘記密碼
評論只代表網(wǎng)友觀點,不代表本站觀點。 請輸入驗證碼: 8795
Copyright(C) 1998-2025 生物器材網(wǎng) 電話:021-64166852;13621656896 E-mail:info@bio-equip.com
主站蜘蛛池模板: 山阴县| 宜兴市| 新营市| 唐山市| 若羌县| 保山市| 耒阳市| 淮北市| 三河市| 禄丰县| 黄石市| 乌拉特中旗| 陈巴尔虎旗| 武安市| 钟山县| 杨浦区| 神木县| 昌邑市| 中西区| 林州市| 历史| 绥阳县| 庐江县| 旬邑县| 九江市| 深水埗区| 商水县| 彰化市| 繁昌县| 丹棱县| 闻喜县| 五莲县| 昌宁县| 综艺| 泗洪县| 东源县| 青海省| 扬中市| 荔浦县| 瓮安县| 延川县|