小鼠真皮成纤维细胞-细胞株/菌种-试剂-生物在线
小鼠真皮成纤维细胞

小鼠真皮成纤维细胞

商家询价

产品名称: 小鼠真皮成纤维细胞

英文名称: MDF

产品编号: XY2300-57

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

小鼠真皮成纤维细胞Cell Specification
Fibroblasts are mesenchymal cells derived from the embryonic mesoderm. They have been
extensively used for a wide range of cellular and molecular studies as they are one of easiest
types of cells to grow in culture. Their durability also makes them amenable to a variety of
manipulations ranging from studies employing gene transfection to microinjection. In general,
fibroblasts secrete a non-rigid extracellular matrix which is rich in type I and/or type III collagen
[1]. There is evidence showing that fibroblasts in different organs are intrinsically different [2].
Dermal fibroblasts switch from a proliferative, migratory phase to a contractile, matrixremodeling
phase during wound healing. In addition, they secrete large quantities of hyaluronan
in response to inflammatory stimuli [3].
MDF from ScienCell Research Laboratories are isolated from postnatal day 2 C57BL/6 mouse
skin. MDF are cryopreserved at passage one and delivered frozen. Each vial contains >5 x 105
cells in 1 ml volume. MDF are characterized by their spindle morphology and
immunofluorescence with antibody specific to fibronectin. MDF are negative for mycoplasma,
bacteria, yeast, and fungi. MDF are guaranteed to further expand for 5 population doublings
under the conditions provided by ScienCell Research Laboratories.
Recommended Medium
It is recommended to use Fibroblast Medium-2 (FM-2, Cat. #2331) for culturing MDF in vitro.
小鼠真皮成纤维细胞Product Use
MDF 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.
Shipping
Dry ice.
References
[1] Gabbiani G, Rungger-Brandle E. (1981) “The fibroblast.” In Glynn LE, Handbook of Inflammation, Vol. 3:
Tissue Repair and Regeneration (pp 1-50). Amsterdam: Elsevier.
[2] Conrad GW, Hart GW, Chen Y. (1977) “Differences in vitro between fibroblast-like cells from cornea, heart,
and skin of embryonic chicks.” J Cell Sci. 26: 119-37.
[3] Stair S, Carlson KW, Shuster S, Wei ET, Stern R. (2002) “Mystixin peptides reduce hyaluronan deposition and
edema formation.” Eur J Pharmacol 450: 291-6.
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 (FM-2, Cat. #2331). Thaw FGS (Cat. #2352), FBS (Cat.
#0010) and P/S solution (Cat. # 0503) at 37 oC. Decontaminate the external surfaces of
medium bottle and medium supplement tubes with 70% ethanol and transfer them to a
sterile field. Gently tilt FGS tube several times to ensure the contents are completely
dissolved before adding to the medium. Aseptically transfer supplement to the basal
medium with a pipette. Rinse the supplement 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
dispense the contents of the vial into the equilibrated, poly-L-lysine-coated culture vessel.
小鼠真皮成纤维细胞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. Change the culture medium the next day to remove residual DMSO and
unattached cells.
Maintaining the culture:
1. Change 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:
1. Subculture when the culture reaches 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.
小鼠真皮成纤维细胞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 animal derived products is potentially biohazardous. Always wear 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 Cult Methods. 11: 191-9.