人口腔成纤维细胞-细胞株/菌种-试剂-生物在线
人口腔成纤维细胞

人口腔成纤维细胞

商家询价

产品名称: 人口腔成纤维细胞

英文名称: HOrF

产品编号: XY2640

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
  • 邮箱 : shxysw02@163.com

人口腔成纤维细胞Cell Specification
Fibroblasts are mesenchymal cells which perform many vital functions during development and
in adulthood. They are responsible for much of the synthesis of extracellular matrix in
connective tissue and play major roles in wound healing. Many diseases are associated with
fibroblasts, either because fibroblasts are implicated in their etiology or because of the fibrosis
that accompanies damage to other cell types. Human oral fibroblasts (HOrF), located in the oral
cavity, have the ability to rapidly repair defects in the oral cavity [1, 2]. HOrF, in contrast to skin
fibroblasts, can more quickly reorganize the extracellular matrix and migrate for wound repair
[1, 2]. Abnormal proliferation of HOrF can lead to the development of oral squamous cell
carcinoma [3]. HOrF are a useful model for elucidating the mechanisms of fibrosis and
developing treatments for oral cancers.
HOrF from ScienCell Research Laboratories are isolated from human oral tissue. HOrF are
cryopreserved at passage one and delivered frozen. Each vial contains >5 x 105
cells in 1 ml
volume. HOrF are characterized by their spindle morphology and immunofluorescence with
antibodies specific to fibronectin. HOrF are negative for HIV-1, HBV, HCV, mycoplasma,
bacteria, yeast and fungi. HOrF are guaranteed to further expand for 15 population doublings
under the conditions provided by ScienCell Research Laboratories.
Recommended Medium
It is recommended to use Fibroblast Medium (FM, Cat. #2301) for the culturing of HOrF in
vitro.
Product Use
HOrF are for research use only. They are not approved for human or animal use, or for
application in in vitro diagnostic procedures.
Storage
Upon receiving, directly and immediately transfer the cells from dry ice to liquid nitrogen and
keep the cells in liquid nitrogen until they are needed for experiments.
人口腔成纤维细胞Shipping
Dry ice.
References
[1] Stephens P, Davies K, Occleston N, Pleass R, Kon C, Daniels J, Khaw P, Thomas D. (2001) “Skin and oral
fibroblasts exhibit phenotypic differences in extracellular matrix reorganization and matrix metalloproteinase
activity.” Br J Dermatol. 144(2): 229-237.
[2] Enoch S, Wall I, Peake M, Davies L, Farrier J, Giles P, Baird D, Kipling D, Price P, Moseley R, Thomas D,
Stephens P. (2009) “Increased oral fibroblast lifespan is telomerase-independent.” J Dent Res. 88(10): 916-921.
[3]Thode C, Jørgensen TG, Dabelsteen E, Mackenzie I, Dabelsteen S. (2011) “Significance of myofibroblasts in oral
squamous cell carcinoma.” J Oral Pathol Me. 40(3): 201-207.
Instructions for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC water bath
and return the cells to culture as quickly as possible with minimal handling!
Initiating the culture:
1. Prepare a poly-L-lysine-coated culture vessel (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, Cat. #0413). Leave the vessel in a 37oC incubator overnight (or for a
minimum of one hour).
2. Prepare complete medium. Decontaminate the external surfaces of medium bottle and
medium supplement tubes with 70% ethanol and transfer them to a sterile field.
Aseptically transfer supplement to the basal medium with a pipette. Rinse the supplement
人口腔成纤维细胞tube with medium to recover the entire volume.
3. Rinse the poly-L-lysine-coated vessel twice with sterile water and then add 15 ml of
complete medium. Leave the vessel in the sterile field and proceed to thaw the
cryopreserved cells.
4. Place the frozen vial in a 37oC water bath. Hold and rotate the vial gently until the
contents completely thaw. Promptly remove the vial from the water bath, wipe it down
with 70% ethanol, and transfer it to the sterile field.
5. Carefully remove the cap without touching the interior threads. Gently resuspend and
人口腔成纤维细胞dispense the contents of the vial into the equilibrated, poly-L-lysine-coated culture vessel.
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 residual DMSO in the culture. It is
also important that cells are plated in poly lysine coated culture vessels to promote cell
attachment.
6. Replace the cap or lid of the culture vessel and gently rock the vessel to distribute the
cells evenly. Loosen cap, if necessary, to allow gas exchange.
7. Return the culture vessel to the incubator.
8. For best results, do not disturb the culture for at least 16 hours after the culture has been
initiated. Refresh culture medium the next day to remove residual DMSO and unattached
cells.
Maintaining the culture:
1. Refresh supplemented culture 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% confluency, change medium every other day until the
culture is approximately 90% confluent.
Subculturing:
人口腔成纤维细胞1. Subculture when the culture reaches 90-95% confluency.
2. Prepare poly-L-lysine-coated culture vessels (2 μg/cm2
) one day before subculture.
3. Warm complete medium, trypsin/EDTA solution (T/E, Cat. #0103), T/E neutralization
solution (TNS, Cat. #0113), and DPBS (Ca++
- and Mg++
-free, Cat. #0303) to room
temperature. We do not recommend warming reagents and medium in a 37oC water bath
prior to use.
4. Rinse the cells with DPBS.
5. Add 8 ml of DPBS and then 2 ml of T/E solution into flask (in the case of a T-75 flask).
Gently rock the flask to ensure complete coverage of cells by T/E solution. Incubate the
flask in a 37oC incubator for 1 to 2 minutes or until cells completely round up. Use a
microscope to monitor the change in cell morphology.
6. During incubation, prepare a 50 ml conical centrifuge tube with 5 ml of fetal bovine
serum (FBS, Cat. #0500).
7. Transfer T/E solution from the flask to the 50 ml centrifuge tube (a small percent of cells
may detach) and continue to incubate the flask at 37oC for another 1 to 2 minutes (no
solution in the flask at this moment).
8. At the end of incubation, gently tap the side of the flask to dislodge cells from the
surface. Check under a microscope to make sure that all cells detach.
9. Add 5 ml of TNS solution to the flask and transfer detached cells to the 50 ml centrifuge
tube. Rinse the flask with another 5 ml of TNS to collect the residual cells.
10. Examine the flask under a microscope for a successful cell harvest by looking at the
number of cells being left behind; there should be less than 5%.
Note: Use ScienCell T/E solution that is optimized to minimize cell damages due to over
trypsinization.
11. Centrifuge the 50 ml centrifuge tube at 1000 rpm for 5 minutes. Resuspend cells in
culture medium.
12. Count and plate cells in a new poly-L-lysine-coated culture vessel with the recommended
cell density.
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
ear gloves and safety glasses when working with 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 WE, Polt S. (1988) “Guidelines to avoid personal contamination by infective agents in research
laboratories that use human tissues.” J Tissue Culture Methods. 11: 191-9.