人脉络丛内皮细胞-细胞株/菌种-试剂-生物在线
人脉络丛内皮细胞

人脉络丛内皮细胞

商家询价

产品名称: 人脉络丛内皮细胞

英文名称: Human Choroid Plexus Endothelial Cells

产品编号: XY1300

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

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人脉络丛内皮细胞Cell Specification
The choroid plexus plays a wide range of roles in brain development, maturation, aging process,
endocrine regulation, and pathogenesis of certain neurodegenerative diseases [1]. The choroid
plexuses consist of a single layer of epithelial cells enclosing a vascular core that together form
the blood-cerebrospinal fluid barrier [2]. The choroidal capillary is a single layer of endothelial
cells interrupted by “pores” which exhibit a diaphragm between the lumen and the interstitial
space. Study shows that choroid plexus endothelial cells express high Glut1 glucose transporter
[3]. The high glucose transport densities in choroids plexus endothelial cells is consistent with
the suggestion that choroids epithelial and endothelial cells provide a metabolic work capability
for maintaining ionic gradients and secretory functions across the blood-cerebrospinal fluid
barriers.
HCPEC from ScienCell Research Laboratories are isolated from human brain. HCPEC are
cryopreserved immediately after purification and delivered frozen. Each vial contains >5 x 105
cells in 1 ml volume. HCPEC are characterized by immunofluorescent method with antibodies to
VWF/Factor VIII, CD31(PCAM) and by uptake of DiI-Ac-LDL. HBMEC are negative for HIV-
1, HBV, HCV, mycoplasma, bacteria, yeast and fungi. HCPEC are guaranteed to further expand
人脉络丛内皮细胞for 15 population doublings in the condition provided by ScienCell Research Laboratories.
Recommended Medium
It is recommended to use Endothelial Cell Medium (ECM, Cat. No. 1001) for the culturing of
HCPEC in vitro.
Product Use
HCPEC are for research use only. It is not approved for human or animal use, or for application
in in vitro diagnostic procedures.
Storage
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
人脉络丛内皮细胞Dry ice.
Reference
[1] Zheng, W. and Zhao, Q. (2002) Establishment and characterization of an immortalized Z310 choroidal epithelial
cell line from murine choroids plexus. Brain Res. 958:371-380.
[2] Thomas, S. A., Bye, A. and Segal, M. B. (2001) Transport Characteristics of the Anti-human Immunodeficiency
Virus Nucleoside Analog, Abacavir, into Brain and Cerebrospinal Fluid. J. Pharmacol. Exp. Ther. 298(3):947-
53.
[3] Cornford, E. M., Hyman, S., Cornford, M. E. and Damian, R. T. (1998) Glut1 glucose transporter in the primate
choroids plexus endothelium. J. Neuropathol. Exp. Neurol. 57:404-411.
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37o
C waterbath
and return them to culture as quickly as possible with minimal handling!
Set up culture after receiving the order:
1. Prepare a fibronectin coated flask (2 μg/cm2
, T-75 flask is recommended). Add 10 ml of
sterile Dulbecco’s phosphate buffered saline (DPBS) to a T-75 flask and then add 150 μl
of fibronectin stock solution (1 mg/ml, Sigma cat. no. F1141). Leave the flask in
incubator overnight.
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.
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
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.
人脉络丛内皮细胞5. Dispense the contents of the vial into the equilibrated, fibronectin-coated flask. A
seeding density of 5,000 cells/cm2
is recommended.
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 endothelial cells are plated in fibronectin coated flask that promotes
cell attachment.
6. Replace the cap or cover of flask, and gently rock the flask to distribute the cells evenly.
Loosen cap if necessary to permit gas exchange.
7. Return the culture vessels to the incubator.
8. For best results, 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, nongranular 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. For subsequent subcultures, change medium 48 hours
after establishing the subculture.
2. Change the medium every other day thereafter, until the culture is approximately 50%
confluent.
3. Once the culture reaches 50% confluence, change medium every 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
) one day before subculture.
3. Warm medium, trypsin/EDTA solution, trypsin neutralization solution, and DPBS to
room temperature. We do not recommend warming the reagents and medium at 37o
C
waterbath prior to use.
4. Rinse the cells with DPBS.
5. Incubate cells with 10 ml of trypsin/EDTA solution (in the case of T-75 flask) until 80%
of cells are rounded up (monitored with microscope). Add 10 ml of trypsin neutralization
solution to the digestion immediately and gently rock the culture vessel.
Note: Use ScienCell Research Laboratories’ trypsin/EDTA solution that is optimized to
minimize the killing of the cells by over trypsinization.
6. Harvest and transfer released cells into a 50 ml centrifuge tube. Rinse the flask with
another 10 ml of growth medium to collect the residue cells. Examine the flask under
microscope to make sure the harvesting is successful by looking at the number of cells
left behind. There should be less than 5%.
7. Centrifuge the harvested cell suspension at 1000 rpm for 5 min and resuspend cells in
growth medium.
8. Count cells and plate them in a new, fibronectin-coated flask with cell density as
recommended.
Caution: Handling human derived products is potentially bioharzadous. Although each cell strain testes negative
for HIV, HBV and HCV DNA, diagnostic tests are not necessarily 100% accurate, therefore, proper precautions
mush 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).