企业档案
会员类型：旗舰版
已获得阿仪网信誉 等级评定
90成长值
(0 -40)基础信誉积累，可浏览访问
(41-90)良好信誉积累，可接洽商谈
(91+ )优质信誉积累，可持续信赖
阿仪网旗舰版会员：9年
工商认证 【已认证】
最后认证时间：
注册号： 【已认证】
法人代表： 【已认证】
参观次数：7950951
手机网站：http://wap.app17.com/c137908/
旗舰版网址：http://www.yajiml.com
技术文章
Mesenchymal Stem Cell Osteogenic Differentiation Medium (MODM)
点击次数：3773 发布时间：2019/5/30 10:07:29
Product Description
Mesenchymal Stem Cell Osteogenic Differentiation Medium (MODM), when used with
Mesenchymal Stem Cell Osteogenic Differentiation Supplements (MODS-A, Cat #7532-A and
MODS-B, Cat #7532-B) ａnd 25 ml of fetal bovine serum, is a complete medium designed for the
optimal osteogenic differentiation of mesenchymal stem cell (MSCs) in vitro. It is a sterile, liquid
medium which contains essential ａnd non-essential amino acids, vitamins, organic ａnd inorganic
compounds, hormones, growth factors, trace minerals, ａnd a low concentration of fetal bovine
serum (5%). The medium is HEPES ａnd bicarbonate buffered ａnd has a pH of 7.4 when
equilibrated in an incubator with an atmosphere of 5% CO2/95% air. The medium is formulated
(quantitatively ａnd qualitatively) to provide an optimally balanced nutritional environment that
selectively promotes osteogenic differentiation of normal human mesenchymal stem cells in vitro.
Components
MODM consists of 500 ml of basal medium, 25 ml of fetal bovine serum (FBS, Cat #0025), 5 ml of
Mesenchymal Stem Cell Osteogenic Differentiation Supplement-A (MODS-A, Cat. #7532-A), 10
ml of Mesenchymal Stem Cell Osteogenic Differentiation Supplement-B (MODS-B, Cat. #7532-B),
and 5 ml of penicillin/streptomycin solution (P/S, Cat. #0503). Note: FBS, MODS-A, MODS-B and
P/S are not pre-mixed in MODM; they must be added separately to make the complete MODM.
Product Use
MODM is for research use only. It is not approved for human or animal use, or for application in in
vitro diagnostic procedures.
Storage
Store the basal medium at 4oC, ａnd the MODS-A, MODS-B, FBS, ａnd the P/S solution at -20oC.
Protect from light.
Shipping
Basal medium: room temperature. Supplements: dry ice.
Prepare for use
Thaw MODS-A, MODS-B, FBS ａnd P/S solution at 37oC. Gently tilt the tubes several times to
ensure the contents are completely mixed before adding to the medium. Spray the medium bottle
and tubes with 70% ethanol, ａnd wipe to remove excess liquid. In a sterile field, remove the caps
without touching the interior threads with fingers. Add MODS-A, MODS-B, FBS ａnd P/S solution
to the medium ａnd mix well. Since several components are light-labile, the medium should not be
exposed to light for extended periods. We do not recommend warming medium in a 37oC water
bath prior to use. When stored in the dark at 4oC, the reconstituted medium is stable for one month.
Caution: If handled improperly, some components of the medium may present a health hazard. Take appropriate
precautions when handling it, including the wearing of protective clothing ａnd eyewear. Dispose of properly.
Instructions for Osteogenic Differentiation
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC water bath ａnd return
them to culture as quickly as possible with minimal handling!
Setup Expansion Culture for Differentiation:
Note: It is recommended to use cells of low passage (≤3 passages) because the efficiency of
differentiation decreases as the number of passages increases.
1. Primary Mesenchymal Stem Cells (MSCs) should be expanded with MSCM (cat # 7501) in
T-25 or T-75 flasks, which have been coated with poly-l-lysine ａnd placed for at least 1 hour
in the 37°C incubator.
2. Change the medium to fresh supplemented medium the next morning after establishing a
culture from cryopreserved cells. For subsequent subcultures, change medium 48 hours after
establishing the subculture.
3. Change the medium every other day thereafter, until the culture is ready for subculture.
4. In general, human MSCs can be subcultured every 3 to 4 days, ａnd rat MSCs can be
passaged every 4 to 6 days
Induction of Osteogenic Differentiation:
1. Prepare a coated 6-well plate or T-25 flask with poly-l-lysine (2μg/cm2
). For a 6-well plate,
add 144 μl of poly-l-lysine (Cat #0403) to 9ml of sterile water. Add 1.5ml of this diluted
poly-l-lysine to each well of the 6-well plate. For a T-25 flask, add 5ml of sterile water
containing 50μl of poly-l-lysine (Cat #0403) to the flask.
Note: Even though some studies suggest that coating tissue culture plastic-ware with vitronectin
and collagen I may promote osteogenic differentiation over non-coated plates, the results obtained
at ScienCell Research Laboratories indicate that ScienCell’s MSC products can easily differentiate
into osteoblasts when plastic-vessels coated with poly-L-lysine are used.
2. Leave the plate or the flask in the 37°C incubator for overnight (or at least 1 hour before
using).
3. The next day, aspirate the poly-l-lysine solution from the wells or flask ａnd rinse the vessels
twice with sterile water, aspirating in between washes.
4. Plate the cell suspension in MSCM at a density of 10,000 cells/cm2
in the coated flask or
plate.
5. Incubate the cells at 37°C in a 5% CO2 humidified incubator for 1-2 days.
Note: Cells should reach 100% confluence before initiating osteogenic induction.
6. When the cells are 100% confluent, carefully replace the MSCM with osteogenic
differentiation medium (MODM, Cat # 7531). This medium change counts at differentiation
day 1.
7. Replace the medium with fresh osteogenic differentiation medium every 3-4 days for T-25
flasks, or every 4-5 days for 6 well plates.
Note: During the induction of osteogenic differentiation, cells can easily peel off from the plates
by the medium changes. Be extremely gentle ａnd careful with the cells during medium changes.
The cells are more easily detached when a smaller vessel area is used. Therefore, 6 well plates
are the smallest multi-well plates recommended for use.
8. After 14-16 days of differentiation, cells can be fixed ａnd stained with Alizarin Red
Solution.
Note: after 6-8 days of incubation with osteogenic differentiation medium, dark brown cell
multilayers will be observed under microscope (4X). White colonies may also be seen with the
naked eye.
Caution: Handling human derived products is potentially biohazardous. Although each cell strain testes negative for
HIV, HBV ａnd HCV DNA, diagnostic tests are not necessarily 100% accurate, therefore, proper precautions mush 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 minimum
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).
A. Human mesenchymal stem cells from bone marrow (HMSC-bm/7500) were cultivated in
expansion medium (Cat. No. 7501) for 16 days. Alizarin Red staining was not detected.
B. HMSC-bm cells were cultivated in osteogenic differentiation medium (Cat. No. 7531) for 16
days. The Alizarin Red staining demonstrates that calcium deposit throughout the culture.
原创作者：上海雅吉生物科技有限公司
