Isolation and cultivation of endothelial progenitor cells (EPCs)

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 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. BM-derived progenitors also contribute to the formation of the microvasculature in allograft arteriosclerotic lesions., EPCs have both physiologic and pathologic roles; thus, they have been widely considered for establishing New therapeutic strategies for the treatmen 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.

Others Finished Drugs

PRODUCT SPECIFICATION NORMAL PACKING
Analgin /Diyron /Metamizol Injection 0.5mg/2ml 2.5g/5ml
Tablet 0.5g
100amps/Box
1000's/Tin
Celecoxib Capsule 200mg 10x10`s/Box
Diclofenac Sodium Inj. 75mg/3Ml 5's/Tray/Box
Ibuprofene Tablet 0.2g 0.4g
Syrup 100mg/5ml 25ml
1000's/Tin
1`s/Box
Indomethacin Capsule 25mg 1000's/Tin
Paracetamol Tablet 0.5g 0.3g
Injection 300mg/2ml
Infusion 1g/100ml
1000's/Box
10ampoules/Box
1`s/Bottle
Piroxicam Capsule 20mg ; 40mg
Injection 20mg/2ml
10`Sx10/Box
100`S/Box
Dexamethason Phosphate Inj. 4mg/Ml 5x5amps/Tray/Box
Hydrocortisone Sodium Succinate Powder Injection 100mg 50vials/Box
Levonorgestrol Tablet 0.75mg 10`s x10/Box
Levonorgestrel +Ethinylestradiol Tablet 0.15mg+0.03mg 21`s/Blister
Prednisolone Tablet 5mg 1000's/Tin
Progesterone Oil Inj. 20mg/Ml ,25mg/Ml 100amps/Box
Testosterone Proplonate Oil Inj.10mg/Ml 25mg/Ml 10'sx10tray/Box
Triamcinolone acetonide Inj. 80mg/2ml 10`s/Box
Adrenine Injection 1mg/Ml 10`s/Box
Atracurium Besylate Powder for inj. 25mg/2.5ml; 50mg/5ml 1`s/Box
Atropine Inj. 1mg/Ml 10'sx10tray/Box
Chlopromazine Hcl Injection 25mg/2ml 100`S/Box
Diazepam Inj. Inj. 20mg/2Ml 10`S/Box
Promethazine Inj. 25mg/2ml 10'sx10tray/Box
Ambroxol HCL syrup 30mg/5ml 100mL 1Bottle/Box
Aminophilline Injection 0.25g 10ml 50's/Box
Salbutamol Aerosol 100mcg/Dose 200Doses 1bottle/Box
Beclomethasone Dipropionate Aerosol 100mcg 200doses 1`s/Box
Xylometazoline Nasal drop 0.1% 10Ml Plastic Vial
Diosmin Tablet Tablet 500mg 10`sx10/Box
Metformini Hcl Tablet 0.25g 1000's/Tin
Orlistat Tablet 120mg 10x10`s/Box
Vecuronium Bromide Powder for Injection 4mg 1Vial/Box

Paclitaxel Injection,Diosmin Tablet Uses,Dexamethasone Phosphate Injection,Vecuronium Bromide Injection

NINGBO VOICE BIOCHEMIC CO. LTD , https://www.pharma-voice.com

Posted on