人小神经胶质细胞
产品名称： 人小神经胶质细胞
英文名称： Human Microglia
产品编号： XY1900
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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人小神经胶质细胞 Cell Specification
Microglia, one of the glial cell types in the CNS, is an important integral component of neuroglial
cell network [1]. They have been observed in the brain parenchyma from the early stage of
development to the mature state. Microglia act as brain macrophages when programmed cell
death occurs during brain development or when the CNS is injured or pathologically damaged.
Microglia can be considered as the main cell in brain immune surveillance, can present antigens
in the molecular context of MHC class II expression to CD-4 positive T cells, are capable of Fcmediated
phagocytosis, and share many common antigens with hemopoietic and tissue
macrophages [2]. Furthermore, there is accumulating evidence that microglia are involved in a
variety of physiological and pathological processes in the brain by interacting with neurons and
other glial cells and through production of biologically active substances such as growth factors,
cytokines, and other factors [3].
HM from ScienCell Research Laboratories are isolated from primary human brain cell culture.
HM are cryopreserved after purification without further expansion and delivered frozen. Each
vial contains >5 x 10^5 cells in 1 ml volume. HM culture in different size of vessels is about
90% confluent and delivered in gel ice box. HM is characterized by immunofluorescent method
with antibody to OX-42 (CD 11b/c). HM is negative for HIV-1, HBV, HCV, mycoplasma,
bacteria, yeast and fungi. HM is guaranteed to further culture in the condition provided by
ScienCell Research Laboratories.
Product Use
人小神经胶质细胞HM are for research use only. They are not approved for human or animal use, or for application
in in vitro diagnostic procedures.
Storage
Transfer cells directly and immediately 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 or gel ice.
Reference
[1] Lee, S. C., Liu, W., Brosnan, C. F. and Dickson, D. W. (1992) Characterization of primary human fetal
dissociated central nervous system cultures with an emphasis on microgia. Laboratory Investigation. 67:465-476.
[2] Fedoroff, S., Zhai, R. and Novak, J. P. (1997) Microglia and astroglia have a common progenitor cell. J.
Neurosci. Res. 50: 477-486.
[3] Stoll, G. and Jander, S. (1999) The role of microglia and macrophages in the pathophysiology of the CNS. Prog.
Neurobiol. 58:233-247.
ScienCell
Research Laboratories
TM
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC waterbath
and return them to culture as quickly as possible with minimal handling!
Set up culture after receiving the ordering:
人小神经胶质细胞1. Place the vial in a 37oC waterbath, hold and rotate the vial gently until the contents are
completely thawed. Remove the vial from the waterbath immediately, wipe it dry, and
transfer it to a sterile field. Rinse the vial with 70% ethanol, and then wipe to remove
excess. Remove the cap, being careful not to touch the interior threads with fingers.
Using 1 ml eppendorf pipet gently resuspends the contents of the vial.
2. Using 1 ml eppendorf pipette gently transfer the contents of the vial into a 15 ml
centrifuge tube; slowly add 10 ml of complete microglia medium to the cells in a way
drop by drop in the first ml. Centrifuge the tube at 1000 rpm for 5 min.
3. Gently resuspend the cell pallet in 7 ml of microglia medium and plate cells in poly-Llysine-coated
culture vessels. A seeding density of >10,000 cells/cm2
(a T-45 flask) is
recommended.
4. 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 microglia are plated in poly-L-lysine coated culture vessels that
promote cell attachment.
5. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly. Loosen
caps if necessary to permit gas exchange.
6. Return the culture vessels to the incubator.
7. 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 health culture of microglia show
characteristic elongated, almost bipolar cell bodies with spine-like processes that often
branch perpendicularly.
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:
Microglia is not supposed to be subcultured since this cell type is terminally differentiated
cells. The following is only for reference in case you need to subculture them.
1. Prepare poly-L-lysine coated cell culture flasks.
2. Warm medium, trypsin/EDTA solution, trypsin neutralization solution, and DPBS to
room temperature. We do not recommend warming the reagents and medium at 37oC
waterbath prior to use.
3. Rinse the cells with DPBS.
4. Incubate cells with 5 ml of trypsin/EDTA solution (in the case of T-45 flask) until 80% of
cells are rounded up (monitored with microscope). Add 5 ml of trypsin neutralization
solution to the digestion immediately and gently rock the culture vessel.
5. 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 15 ml centrifuge tube. Rinse the flask with
another 3 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.
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.
人小神经胶质细胞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).