人毛囊外根鞘细胞-细胞株/菌种-试剂-生物在线
人毛囊外根鞘细胞

人毛囊外根鞘细胞

商家询价

产品名称: 人毛囊外根鞘细胞

英文名称: Human Hair Follicular Outer Root Sheath Cells

产品编号: XY2420

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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  • 邮编 : 200612
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人毛囊外根鞘细胞Cell Specification
The outer root sheath (ORS) of the hair follicle surrounds the hair fiber and inner root sheath. It
is distinct from other epidermal components being continuous with the surface epidermis. The
ORS consists of several layers of cells that can be identified with unique ultrastructural
properties; plays a part in certain functions of the hair follicle, including acting as the sensory
organ and immunologic sentinel of the skin. Immunohistochemical staining indicates that the
ORS contains nestin-expressing progenitor or stem cells [1]. Human outer root sheath cells
represent a repeatedly available source of keratinocytes which can be easily and extensively
expanded in culture [2].
HHFORSC from ScienCell Research Laboratories are isolated from human hair follicle outer
root sheath. HHFORSC are cryopreserved at primary culture and delivered frozen. Each vial
contains >5 x 105
cells in 1 ml volume. HHFORSC are characterized by their mesenchymal cell
morphology and immunofluorescent method with antibody to fibronectin and CD105.
HHFORSC are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi.
HHFORSC are guaranteed to further expand for 10 population doublings at the condition
provided by ScienCell Research Laboratories.
Recommended Medium
It is recommended to use Mesenchymal Stem Cell Medium (MSCM, Cat. No. 7501) for the
culturing of HHFORSC in vitro.
Product Use
HHFORSC are for research use only. It is not approved for human or animal use, or for
人毛囊外根鞘细胞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
Dry ice.
Reference
[1] Li, L., Mignone, J., Yang, M., Matic, M., Penman, S., Enikolopov, G. and Moffman, R. (2003) Nestin
expression in hair follicle sheath progenitor cells. PNAS 100:9958-9961.
[2]. Limat, A., Breitkreutz, D., Thiekoetter, G., Klein, C.E., Braathen, L.R., Hunziker, T., Fusenig, N.E. (1995)
Formation of a regular neo-epidermis by cultured human outer root sheath cells grafted on nude mice.
Transplantation. 59:1032-8.
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 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
人毛囊外根鞘细胞mg/ml, ScienCell cat. no. 0413). Leave the flask in incubator overnight (minimum one
hour at 37oC 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 37oC 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
also important that cells are plated in poly lysine coated flask that promotes cell
attachment and growth.
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.
Maintenance of Culture:
人毛囊外根鞘细胞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.
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 poly-L-lysine coated cell culture flasks (2 μg/cm2
).
3. Warm medium, trypsin/EDTA solution (T/E, cat. no. 0103), trypsin neutralization
solution (TNS, cat. no. 0113), and DPBS (Ca++ and Mg++ free, cat. no. 0303) to room
temperature. We do not recommend warming the reagents and medium at 37oC
waterbath prior to use.
4. Rinse the cells with DPBS.
5. Add 8 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 37oC incubator for 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 37oC for 1 or 2 minutes more (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%.
Note: Use ScienCell Research Laboratories’ trypsin/EDTA solution that is optimized to
minimize the killing of the cells by over trypsinization.
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, poly-L-lysine coated flask with cell density as
recommended.
Caution: Handling human derived products is potentially biohazardous. Although each cell strain tests negative for
HIV, HBV and HCV DNA, diagnostic tests are not necessarily 100% accurate, therefore, proper precautions must
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
inimum 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).