人脊髓星形胶质细胞-细胞株/菌种-试剂-生物在线
人脊髓星形胶质细胞

人脊髓星形胶质细胞

商家询价

产品名称: 人脊髓星形胶质细胞

英文名称: Human Astrocytes-spinal cord

产品编号: XY1820

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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人脊髓星形胶质细胞Cell Specification
Astrocytes make up the majority of the cells in the mammalian brain. They are the most variable
in type, most intimately associated with all parts of neurons, and thus most functionally
interesting in their relationships with neurons [1]. They provide structural, trophic, and metabolic
support to neurons and modulate synaptic activity. Impairment of these astrocyte functions
during stroke and other insults can critically influence neuron survival. Furthermore, astrocytes
have been implicated in the pathological processes of many neurological diseases [2]. Long-term
recovery after brain injury, through neurite outgrowth, synaptic plasticity, or neuron
regeneration, is influenced by astrocyte surface molecule expression and trophic factor release
[3]. In addition, the death or survival of astrocytes themselves may affect the ultimate clinical
outcome. Recognition of the importance of astrocytes in nervous system functioning is
increasing, specifically regarding the modulation of neural activity. Much of what we have
learned about astrocytes is from the in vitro studies and astrocyte cultures are continuing to
provide a useful tool in exploring the diverse property of these cells.
HA-sp from ScienCell Research Laboratories are isolated from human spinal cord tissue. HA-sp
are cryopreserved at passage one and delivered frozen. Each vial contains >1 x 106
cells in 1 ml
volume. HA-sp are characterized by immunofluorescent method with antibody to GFAP. HA-sp
人脊髓星形胶质细胞are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi. HA-sp are
guaranteed to further expand for 15 population doublings at the conditions provided by ScienCell
Research Laboratories.
Product Use
HA-sp 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 is needed for experiments.
Shipping
Dry ice.
Reference
[1] G. I. Hatton (2002) Glial-neuronal interactions in the mammalian brain. Adv. in Physiol. Edu. 26:225-237.
[2] Van der Laan, L. J. W., De Groot, C. J. A., Elices, M. J. and Dijkstran, C. D. (1997) Extracellular matrix proteins
expressed by human adult astrocytes in vivo and in vitro: an astrocyte surface protein containing the CS1 domain
contributes to binding of lymphoblasts. J. Neurosci. Res. 50:539-548.
[3] Chen Y., and Swanson, R. A. (2003) Astrocytes and brain injury. J. Cereb. Blood Flow Metab. 23:137-149.
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 poly-L-lysine coated flask (2 μg/cm2
, T-75 flask is recommended). Add 10 ml
of sterile water to a T-75 flask and then add 15 μl of poly-L-lysine stock solution (10
mg/ml, ScienCell cat. no. 0413). Leave flask in incubator overnight (minimum one hour
at 37o
C incubator).
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. Rinse the poly-L-lysine coated flask with sterile water twice and add 20 ml of complete
medium to the flask. Leave the flask in the hood and go to thaw the cells.
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.
5. Dispense the contents of the vial into the equilibrated, poly-L-lysine coated culture
vessels. 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
人脊髓星形胶质细胞also important that astrocytes are plated in poly-L-lysine coated culture vessels that
promote cell attachment.
6. Replace the cap or cover of flask, 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 stellar
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 80% confluent.
Subculture:
1. Subculture the cells when they are over 90% confluent.
2. Prepare poly-L-lysine coated cell culture flasks (2 μg/cm2
).
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. Transfer the 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, poly-L-lysine coated flask with cell density as
recommended.
人脊髓星形胶质细胞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).