人脐静脉内皮细胞-细胞株/菌种-试剂-生物在线
人脐静脉内皮细胞

人脐静脉内皮细胞

商家询价

产品名称: 人脐静脉内皮细胞

英文名称: Human Umbilical Vein Endothelial Cells

产品编号: XY8000

产品价格: 0

产品产地: 中国/美国

品牌商标: XYbscience

更新时间: 2023-08-17T09:55:27

使用范围: null

上海信裕生物科技有限公司
  • 联系人 : 徐经理
  • 地址 : 上海市闵行莘庄工业区春东路508号A1-2F
  • 邮编 : 200612
  • 所在区域 : 上海
  • 电话 : 152****8802
  • 传真 : 021-37680378
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人脐静脉内皮细胞Cell Specification
The vascular endothelial cells contribute to the maintenance of vascular homeostasis. They
synthesize and secrete activators as well as inhibitors of both the coagulation system and the
fibrinolysis system in addition to mediators that influence the adhesion and aggregation of blood
platelets. Endothelial cells also release molecules that control cell proliferation and modulate
vessel wall tone. Many of the endothelial processes can be studied in vitro using cultured cells,
and human umbilical vein endothelial cells (HUVEC) are the most commonly used cell type for
such studies. Except for the common endothelial cell features such as “cobblestone”
morphology, positive staining for Factors VIII and the ability to take up acetylated low-density
lipoprotein [1], HUVEC stein positively with CD-31 [2]. Cells pretreated with IL-1 or TNFalpha
also selectively express E-selectin [3] and VCAM [4].
HUVEC from ScienCell Research Laboratories are isolated from human umbilical veins.
人脐静脉内皮细胞HUVEC are cryopreserved at passage one and delivered frozen. Each vial contains >5 x 105
Recommended Medium
cells in 1 ml volume. HUVEC are characterized by immunofluorescent method with antibodies
to vWF/Factor VIII and CD31 and by uptake of DiI-Ac-LDL. HUVEC are negative for HIV-1,
HBV, HCV, mycoplasma, bacteria, yeast and fungi. HUVEC are guaranteed to further expand
for 15 population doublings at the conditions provided by ScienCell Research Laboratories.
It is recommended to use endothelial cell medium (ECM, Cat. No. 1001) for the culturing of
HUVEC in vitro.
Product Use
HUVEC are for research use only
Storage
. It is not approved for human or animal use, or for application
in in vitro diagnostic procedures.
Directly and immediately transfer cells from dry ice to liquid nitrogen upon receiving and keep
the cells in liquid nitrogen until cell culture needed for experiments.
Shipping
Dry ice.
Reference
[1] Morgan, D. M. L. (1996) Isolation and culture of human umbilical vein endothelial cells. In “Human Cell
Culture Protocols” (Gareth E. Jones, eds.) Humana Press, Totowa, New Jersey, pp.104-109.
[2] Newman, P. J. et al., (1990) PECAM-1 (CD31) cloning and relation to adhesion molecules of the
immunoglobulin gene superfamily. Science 247:1219-1222.
[3] Bevilaqua, M. P. et al., (1989) Endothelial leukocyte adhesion molecule 1: an inducible receptor for neutrophils
related to complement regulatory proteins and lectins. Science 243:1160-1165.
[4] Osborn, L. et al., (1989) Direct cloning of vascular cell adhesion molecule 1, a cytokine induced endothelial
protein that binds to lymphocytes. Cell 59:1203-1211.
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37o
and return them to culture as quickly as possible with minimal handling!
C waterbath
Set up culture after receiving the order:
1. Prepare a fibronectin coated flask (2 μg/cm2
, T-75 flask is recommended). Add 5 ml of
人脐静脉内皮细胞sterile Dulbecco’s phosphate buffered saline, Ca++ and Mg++
2. Prepare complete medium: decontaminate the external surfaces of medium and medium
supplements with 70% ethanol and transfer them to sterile field. Aseptically open each
supplement tube and add them to the basal medium with a pipette. Rinse each tube with
medium to recover the entire volume.
free (ScienCell, Cat. No.
0303) to a T-75 flask and then add 150 μl of fibronectin stock solution (ScienCell, Cat.
No. 8248). Leave the flask in incubator overnight.
3. Aspirate fibronectin solution and add 20 ml of complete medium to the flask. Leave the
flask in the hood and go to thaw the cells. The fibronectin solution can be used twice.
4. Place the vial in a 37o
5. Dispense the contents of the vial into the equilibrated, fibronectin coated culture vessels.
A seeding density of 5,000 cells/cm
C waterbath, hold and rotate the vial gently until the contents are
completely thawed. Remove the vial from the waterbath immediately, wipe it dry, rinse
the vial with 70% ethanol and transfer it to a sterile field. Remove the cap, being careful
not to touch the interior threads with fingers. Using a 1 ml eppendorf pipette gently resuspend
the contents of the vial.
2
Note: Dilution and centrifugation of cells after thawing are not recommended since these
actions are more harmful to the cells than the effect of DMSO residue in the culture. It is
also important that cells are plated in fibronectin coated flask that promotes cell
attachment and growth.
is recommended.
6. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly. Loosen
cap if necessary to permit gas exchange.
7. Return the culture vessels to the incubator.
8. For best result, do not disturb the culture for at least 16 hours after the culture has been
initiated. Change the growth medium the next day to remove the residual DMSO and
unattached cells, then every other day thereafter. A healthy culture will display
cobblestone or spindle shaped morphology, non-granular cytoplasm and the cell number
will be double after two to three days in culture.
Maintenance of Culture:
1. Change the medium to fresh supplemented medium the next morning after establishing a
culture from cryopreserved cells.
2. Change the medium every three days thereafter, until the culture is approximately 70%
confluent.
人脐静脉内皮细胞3. Once the culture reaches 70% confluence, change medium every other day until the
culture is approximately 90% confluent.
Subculture:
1. Subculture the cells when they are over 90% confluent.
2. Prepare fibronectin coated flasks (2 μg/cm2
3. Warm medium, trypsin/EDTA solution (T/E, cat. no. 0103), trypsin neutralization
solution (TNS, cat. no. 0113), and DPBS to room temperature. We do not recommend
warming the reagents and medium at 37
) one day before subculture.
o
4. Rinse the cells with DPBS.
C waterbath prior to use.
5. Add 10 ml of DPBS first and then 2 ml of trypsin/EDTA solution into flask (in the case
of T-75 flask); gently rock the flask to make sure cells are covered by trypsin/EDTA
solution; incubate the flask at 37o
C incubator for 1 to 2 minutes or until cells are
completely rounded up (monitored with inverted microscope). During incubation, prepare
a 50 ml conical centrifuge tube with 5 ml of fetal bovine serum (FBS, cat. no. 0500);
transfer trypsin/EDTA solution from the flask to the 50 ml centrifuge tube (a few percent
of cells may detached); continue incubate the flask at 37o
Note: Use ScienCell Research Laboratories’ trypsin/EDTA solution that is optimized to
minimize the killing of the cells by over trypsinization.
C for 1 minutes (no solution in
the flask at this moment); at the end of trypsinization, one hand hold one side of flask and
the other hand gently tap the other side of the flask to detach cells from attachment; check
the flask under inverted microscope to make sure all cells are detached, add 5 ml of
trypsin neutralization solution to the flask and transfer detached cells to the 50 ml
centrifuge tube; add another 5 ml of TNS to harvest the residue cells and transfer it to the
50 ml centrifuge tube. Examine the flask under inverted microscope to make sure the cell
harvesting is successful by looking at the number of cells left behind. There should be
less than 5%.
6. Centrifuge the 50 ml centrifuge tube (harvested cell suspension) at 1000 rpm (Beckman
Coulter Allegra 6R centrifuge or similar) for 5 min; re-suspend cells in growth medium.
7. Count cells and plate cells in a new, fibronectin coated flask with cell density as
recommended.
Caution: Handling human derived products is potentially biohaza dous. Although each cell strain tests negative for
HIV, HBV and HCV DNA, diagnostic tests are not necessarily 100% accurate, therefore, proper precautions mus
be taken to avoid inadvertent exposure. Always wear gloves and safety glasses when working these materials. Never
mouth pipette. We recommend following the universal procedures for handling products of human origin as the
minimum precaution against contamination [1].
[1]. Grizzle, W. E., and Polt, S. S. (1988) Guidelines to avoid personal contamination by infective agents in research
laboratories that use human tissues. J Tissue Culture Methods. 11(4).