Newsletter   |   Register   |   Log In   |   Cart(0)   
Shopping Cart
0 items
 

NEWS


Cell Biologics' products can be purchased through Fisher Scientic by searching Cell Biologics' product catalog number or key words...

  • Medium Promotion, Buy 3 Bottles, get the 4th Free on Feb. 2018.
  • Promo Code: CB0218.

Meet Us

  • AACR April 14-18, 2018 - Chicago
  • Experimental Biology April 22-24, 2018 - San Diego

PROMOTIONS


  • Get Free Gelatin-Based Coating Solution per primary cell order, which can be used for most primary cell culture from Cell Biologics through 2018.

NEW PRODUCTS


Cynomolgus Monkey Primary Dermal Fibroblasts

Cynomolgus Monkey Primary Dermal Fibroblasts
Click to enlarge
Price: $0.00
Cat. #: MK-6067

Available Options:
Sequence:
Purity:
Primary Cells:
Qty: Buy Quote

Related Documents:

Cynomolgus Monkey Primary Dermal Fibroblasts

Catalog No. MK-6067

Suggested Medium:  Catalog No. M2267 Fibroblast Medium /w Kit (500 ml)

 

Product Description

Monkey Primary Dermal Fibroblasts from Cell Biologics are isolated from tissue of Cynomolgus Monkey. Monkey Primary Dermal Fibroblasts are grown in T25 tissue culture flasks pre-coated with gelatin-based solution for 0.5 hour and incubated in Cell Biologics Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded.  Prior to shipping, cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x106 cells per ml and is delivered frozen. Monkey Primary Dermal Fibroblasts are negative for bacteria, yeast, fungi, and mycoplasma. Cells can be expanded for 3-6 passages at a split ratio of 1:3 under the cell culture conditions specified by Cell Biologics. Repeated freezing and thawing of cells is not recommended.

Laboratory Applications

Standard biochemical procedures performed with cell cultures include the assay of cell to cell interaction, RT-PCR, Western blotting, immunoprecipitation, immunofluorescent staining, flow cytometry or generating cell derivatives for desired research applications.

Storage of Cell Biologics Products 

Cell Biologics will ship frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen (-180°C) until ready for experimental use. Live-cell shipment is also available on request.

Authorized Uses of Cell Biologics’ Products 

Monkey Primary Dermal Fibroblasts from Cell Biologics are distributed for research purposes only. Our products are not authorized for human use, for in vitro diagnostic procedures or for therapeutic procedures. Transfer or resale of any Cell Biologics’ cells or products from the purchaser to other markets, organizations or individuals is prohibited by Cell Biologics without the company’s written consent. Cell Biologics’Terms and Conditions must be accepted before submitting an order.

Disclaimer

Investigators should handle the cells that they receive from Cell Biologics with caution and treat all animal cells as potential pathogens, since no test procedure can completely guarantee the absence of infectious agents.

Warranty and Liability

CELL BIOLOGICS’ guarantee applies only to your purchase of CELL BIOLOGICS' cells with CELL BIOLOGICS’ Media and Coating Solution for appropriate cell culture within 35 days from the date of product delivery.

 

Primary Cell Culture Protocol

>>>>>>>>>>>>>>>>>>>>>>>> 

All cell culture procedures must be conducted in a bio-safety cabinet. Any and all media, supplements, and reagents must be sterilized by filtration through a 0.2 µm filter. Use aseptic technique to prevent microbial contamination. Cryo-preserved cells must be stored in liquid nitrogen or seeded immediately upon arrival.

Medium:

Review the information provided on the Cell Biologics website about appropriate culture media (e.g. serum and other supplements). Use pre-warmed (37°C) cell culture media (30-50 ML) to recover cryo-preserved cells and when changing media or splitting cells.

Coating of flasks or dishes:

Coat sterile culture dishes or flasks with Gelatin-Based Coating Solution (Cell Biologics, Catalog No. 6950) for 10-30 min and then aspirate the excess solution before seeding cells.

Cell recovery from cryovial:

  • Quickly thaw cells in cryo-vial by incubating them in a 37°C water bath for <1 min until there is just a small bit of ice left in the vial.
  • Promptly remove the vial and wipe it down with 70% ethanol.
  • Transfer cells from the vial to a sterile centrifuge tube. Add 8-10 ml of pre-warmed Cell Biologics Cell Culture Medium. Flush the vial with an additional 0.5-1 ml of medium to ensure complete transfer of cells to the centrifuge tube.
  • Centrifuge cells at 200 g for 5 minutes.
  • Aspirate the supernatant and resuspend the cell pellet in 6 ml of Cell Biologics’Cell Culture Growth Medium.
  • Add resuspended cells into a T25 flask pre-coated with Gelatin-Based Coating Solution (Cell Biologics, Catalog No. 6950).
  • Place the T25 flask in a humidified, 5%-CO2 incubator at 37°C.
  • Change culture media the following day to remove non-adherent cells and replenish nutrients.
  • Change cell culture medium everyday when cells are >60% confluent.
  • Cells should be checked daily under a microscope to verify appropriate cell morphology.

Expansion of cultured mouse primary cells:

  • Flush the adherent layer with a 5 ml sterile pipette 3 times to dislodge loosely attached cells.
  • Remove and discard the cell culture media from the flask.
  • Wash adherent cells 2 times with 10 ml of sterile PBS (1X) without calcium and magnesium to remove nonadherent cells or fraction.
  • Remove and discard the wash solution from the flask.
  • Incubate cells with warm (37°C) 0.05% Trypsin-EDTA (1X) solution (Invitrogen catalog number 25300) for 1 minutes. Use 2.0 ml of 0.05% Trypsin-EDTA solution when collecting cells from T75 flasks, and 1.0 ml when using T25 flasks. As soon as cells have detached (the flask may require a few firm gentle taps), add 8-10 ml of Cell Biologics Cell Culture Medium supplemented with 5-10 % FBS to a T25 or T75 flask (the FBS will neutralize the trypsin).
  • Plate cells in fresh flasks or plates precoated with Gelatin-Based Coating Solution in a humidified, 5%-CO2 incubator at 37°C. 
  • Change culture media the following day to remove non-adherent cells and replenish nutrients.
  • Cells should be checked daily under a microscopy to verify appropriate cell morphology.
  • Change culture medium every 24-48 hours. Please note that the medium should be changed every day when cells are >60% confluent to remove non-adherent cells and replenish nutrients.
  • Pre-wash cells with 1X PBS 2 times whenever replacing the medium.

We recommend splitting primary cells at the follow ratio:

  • The recommended split ratio for primary murine cells is 1:2 or 1:3.
  • A confluent monolayer of primary cells grown in a T75 flask may be expanded on a 6-well plate ready for use in experiments under the cell culture conditions specified by Cell Biologics.

Procedure for Freezing Cells

Materials:

  • 1X Phosphate Buffered Saline (PBS-1X)
  • 0.05% Trypsin-EDTA (1X) solution (Invitrogen catalog number 25300)
  • Tissue Culture Media
  • Cold Freezing Media (10% dimethyl sulfoxide (DMSO), 20% FBS and 70% culture medium)
  • Labeled Cryovials
  • Confluent cells

         ----------------------------------

  • Flush the adherent cell layer with a 5 ml sterile pipette 3-5 times to dislodge loosely attached cells.
  • Remove and discard the cell culture media from the flask.
  • Wash adherent cells 2-3 times with 10 ml of sterile PBS (1X) without calcium and magnesium to Remove nonadherent cells or fraction.
  • Remove and discard the wash solution from the flask.
  • Incubate cells with warm (37°C) Trypsin-EDTA (1X) solution for 1 minutes. Use 2.0-3.0 ml of 0.05% Trypsin-EDTA solution when collecting cells from T75 flasks, and 1.0-1.5 ml when using T25 flasks. As soon as cells have detached (the flask may require a few firm gentle taps), add 10 ml of Cell Culture Medium supplemented with 5-10 % FBS to the flask (the FBS will neutralize the trypsin).
  • Centrifuge the cell suspension at 200 g for 5 minutes.
  • Remove supernatant with sterile Pasteur pipette.
  • Quickly re-suspend pellet by adding 1 ml freezing media per vial to be frozen.
  • Place vials in Nalgene "Mr. Frosty" freezing container containing100% isopropyl alcohol at -70-80 °C for 24 h.
  • Transfer vials to liquid N2 tank for indefinite storage.

We recommend freezing primary cells at the follow ratio:

  • A confluent primary cells grown in a T75 flask may be frozen in 3 cryovials.
  • A confluent primary cells grown in a T25 flask may be frozen in 1 cryovial.
Write Review
Your Name:


Your Review: Note: HTML is not translated!

Rating: Bad            Good

Enter the code in the box below:

There are no additional images for this product.

Have you ever published using Cynomolgus Monkey Primary Dermal Fibroblasts? Submit your publication and earn rewards points which can be used for merchandise & discounts. Please include the product used, your name, email, publication title, author(s), PUBMED ID, Journal and issue in your submission.

No Publications Yet.
Related Products
Cat. # Name
6912 Fetal Bovine Serum Buy Quote
6914 Trypsin/EDTA 0.25% Solution Buy Quote
6915 Trypsin/EDTA 0.05% Solution Buy Quote
6916 Cell Culture Freezing Medium - ready to use Buy Quote
6917 Penicillin/Streptomycin Solution (1:100) Buy Quote
6920 Antibiotic/Antimycotic Solution (1:100) Buy Quote
6950 Gelatin-Based Coating Solution- ready to use Buy Quote
6953 Collagen Type I, Rat Tail Solution - ready to use Buy Quote
B129-7067 B129 Mouse Primary Dermal Fibroblasts- Adult Buy Quote
C57-6067 C57BL/6 Mouse Primary Dermal Fibroblasts - Adult Buy Quote
L6811 Tissue Lysis Buffer (1X) Buy Quote
L6822 Cell Lysis Buffer (1X) Buy Quote
M2267 Complete Fibroblast Medium /w Kit – 500 ML Buy Quote
M2267PF Fibroblast Medium (No Phenol Red) /w Kit – 500 ML Buy Quote
M2267SF Serum-Free Fibroblast Medium /w Kit – 500 ML Buy Quote
M2267b Fibroblast Basal Medium – 500 ML Buy Quote
RA-6067 Rat Primary Dermal Fibroblasts Buy Quote