Human Fibroblast Serum-Free Medium for cell proliferation

BBARL, Inc.

Xyltech™ Growth H-Fbro
Catalog Number (BBARL/NIPRO):

10311/87-282

500 mL1. Product features
This product is a synthetic culture medium suitable for cell growth of normal human fibroblasts. When usedin combination with Xyltech™ H-Fbro-01, human fibroblast proliferation can be controlled as needed.
This product is serum-free and contains no human or other animal-derived ingredients. If necessary,
human serum (HS), fetal bovine serum (FBS), or serum substitute reagents can be added for cell culture.
2. Precautions for use
Xyltech™ Growth H-Fbro does not contain substances that neutralize trypsin activity. When subculturing
cells with trypsin, it is strongly recommended that the trypsin activity be sufficiently neutralized with atrypsin inhibitor. Dilution washing alone does not completely remove trypsin activity and the remaining
protease activity will reduce subsequent cell growth. This product is a research reagent. It cannot beused for human or animal treatment or diagnostic purposes.
3. Storage
Store Xyltech™ Growth H-Fbro in a cool, dark place (2-8°C).

Do not freeze the medium to avoid

deterioration of some active ingredients.
4. Example of cell culture protocol for normal human fibroblast culture using Xyltech™ GrowthH-Fbro
4-1. Cells and reagents・Normal human fibroblasts (100 mm-dish)・Xyltech™ Growth H-Fbro (growth medium)*This product・Artificial serum (Xf) or Artificial serum (Af)(NIPRO CAT. #: 87-081/87-082)

・r-TE (r-Trypsin/EDTA Solution)(NIPRO CAT. #: 87-974)

・s-TI (Synthetic Trypsin Inhibitor Solution)(NIPRO CAT. #: 87-975)

・D-PBS(-)*We recommend adding 1% artificial serum (Xf) or artificial serum (Af) for use with Xyltech™ Growth HFbro.
4-2. Cell culture of normal human fibroblasts for growth
1. Warm the culture medium, D-PBS (-), r-TE, and s-TI in a 37°C water bath.
2. Remove the culture supernatant of selected normal human fibroblasts that have reached around
80% confluence (sub confluence).
3. Rinse the cell layer with 5 mL of D-PBS (-).

4. Add 0.5 mL of r-TE and incubate at 37°C for approximately 2 minutes.
5. Add 0.5 mL of s-TI, mix well, gently pipette up and down several times, collect cells from the dish,and centrifuge at 1,000 rpm, for 5 minutes.
6. Aspirate supernatant and add the appropriate amount of Xyltech™ Growth H-Fbro medium toresuspend the cells and seed into a new tissue culture dish.
7. The cells become confluent within 2-4 days when cultured in Xyltech™ Growth H-Fbro medium.
Start subculture and/or experiments with the cells.
8. For experiments requiring reduced growth rate, the cells can be cultured from 3 days to 1 week inXyltech™ H-Fbro-01 growth suppressive medium (See protocol for suppressive growth control using
Xyltech™ H-Fbro-01, BBARL CAT. #: 10301).
4-3. Phase contrast microscope images of normal human fibroblasts cultured with Xyltech™
Growth H-Fbro or Xyltech™ H-Fbro-01

Bars=200 µm

Fig. 1 Normal human fibroblasts cultured
with Xyltech™ Growth H-Fbro for 3 days.

Fig. 2 Normal human fibroblasts cultured
with Xyltech™ H-Fbro-01 (growthsuppressive medium) for 3 days.

*The protocol is based on experimental results. It may be necessary to adjust seeding density, andpassage timing according to the cells. This protocol is intended for research purposes only.

5. For Inquiries about product
Bourbon Biomedical Advanced Research Laboratories, Inc. (BBARL, Inc.)
1-3-1, Ekimae, Kashiwazaki-city, Niigata,
TEL: +81-257-23-2769 E-mail: support@bourbon-barl.co.jp
Copyright©2020 Bourbon Biomedical Advanced Research Laboratories, Inc., All rights reserved.