人前脂肪细胞--内脏-细胞株/菌种-试剂-生物在线
人前脂肪细胞--内脏

人前脂肪细胞--内脏

商家询价

产品名称: 人前脂肪细胞--内脏

英文名称: Human Preadipocytes-visceral

产品编号: XY7210

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

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人前脂肪细胞--内脏Cell Specification
Adipocytes play an important role in energy storage and metabolism. Adipocyte differentiation is
a developmental process that is critical for metabolic homeostasis and nutrient signaling. It is
controlled by complex actions involving gene expression and signal transduction [1].
Preadipocytes are present throughout adult life in adipose tissue and can proliferate and
differentiate into mature adipocytes according to the energy balance [2]. The proliferation and
differentiation of these preadipocytes contribute to increases in adipose tissue mass. In vitro
study indicates that different tissue-derived preadipocytes exhibit differently in lipid
accumulation, adipogenic transcription factor expression, and TNFα-induced
HPA-v from ScienCell Research Laboratories are isolated from human visceral fat tissue. HPA-v
are cryopreserved at primary culture and delivered frozen. Each vial contains >1 x 10
apoptosis [3]. It has
also been demonstrated that there is a close relationship between adipocyte differentiation and
many physiological and pathological processes including fat metabolism, energy balance,
obesity, diabetes, hyperlipidemia and breast cancer.
6
Recommended Medium
cells in 1
ml volume. HPA-v are characterized by immunofluorescent method with antibodies to CD44,
人前脂肪细胞--内脏CD90 and lipid staining after differentiation. HPA-v are negative for HIV-1, HBV, HCV,
mycoplasma, bacteria, yeast and fungi. HPA-v are guaranteed to further culture at the conditions
provided by ScienCell Research Laboratories.
It is recommended to use Preadipocyte Medium (PAM, Cat. No. 7211) for the culturing of HPAv
in vitro. Preadipocyte Differentiation Medium (PADM, Catalog No. 7221) can be use for in
vitro differentiation of preadipocytes into mature adipocytes, and follows by Adipocyte Medium
(AdM, Catalog No. 7201) which maintains mature adipocytes after differentiation.
Product Use
HPA-v are for research use only. It is not approved for human or animal use, or for application in
in vitro diagnostic procedures.
Storage
Directly and immediately transfer cells from dry ice to liquid nitrogen upon receiving and keep
the cells in liquid nitrogen until cell culture is needed for experiments.
Shipping
Dry ice.
Reference
[1] Tominaga, K., Johmura, Y., Nishizuka, M., Imagawa, M. (2004) Fad24, a mammalian homolog of Noc3p, is a
positive regulator in adipocyte differentiation. J Cell Sci. 117(Pt 25):6217-26.
[2] Reue, K., Glueck, S. B. (2001) Accumulating evidence for differences during preadipocyte development: Focus
on "Differential gene expression in white and brown preadipocytes". Physiol Genomics. 7(1):1-2.
[3] Tchkonia, T. et al. (2005) Abundance of Two Human Preadipocyte Subtypes with Distinct Capacities for
Replication, Adipogenesis, and Apoptosis Varies among Fat Depots. Am J Physiol Endocrinol Metab.
288(1):E267-77.
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37o
and return them to culture as quickly as possible with minimal handling!
C waterbath
Set up culture after receiving the order:
1. Prepare a poly-L-lysine coated flask (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, ScienCell cat. no. 0413). Leave the flask in incubator overnight (minimum one
hour at 37o
2. Prepare complete medium: decontaminate the external surfaces of medium and medium
supplements with 70% ethanol and transfer them to sterile field. Aseptically open each
supplement tube and add them to the basal medium with a pipette. Rinse each tube with
medium to recover the entire volume.
C incubator).
3. Rinse the poly-L-lysine coated flask with sterile water twice and add 20 ml of complete
medium to the flask. Leave the flask in the hood and go to thaw the cells.
4. Place the vial in a 37o
5. Dispense the contents of the vial into the equilibrated, poly-L-lysine coated culture
vessels. A seeding density of 5,000 cells/cm
C waterbath, hold and rotate the vial gently until the contents are
completely thawed. Remove the vial from the waterbath immediately, wipe it dry, rinse
the vial with 70% ethanol and transfer it to a sterile field. Remove the cap, being careful
not to touch the interior threads with fingers. Using a 1 ml eppendorf pipette gently resuspend
the contents of the vial.
2
Note: Dilution and centrifugation of cells after thawing are not recommended since these
actions are more harmful to the cells than the effect of DMSO residue in the culture. It is
also important that cells are plated in poly lysine coated flask that promotes cell
attachment and growth.
is recommended.
6. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly. Loosen
cap if necessary to permit gas exchange.
7. Return the culture vessels to the incubator.
8. For best result, do not disturb the culture for at least 16 hours after the culture has been
initiated. Change the growth medium the next day to remove the residual DMSO and
unattached cells, then every other day thereafter.
Maintenance of Culture:
1. Change the medium to fresh supplemented 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% confluence, change medium every other day until the
culture is approximately 90% confluent.
Subculture:
1. Subculture the cells when they are over 90% confluent.
2. Prepare poly-L-lysine coated cell culture flasks (2 μg/cm2
3. Warm medium, trypsin/EDTA solution (T/E, cat. no. 0103), trypsin neutralization
solution (TNS, cat. no. 0113), and DPBS (Ca
).
++ and Mg++ free, cat. no. 0303) to room
temperature. We do not recommend warming the reagents and medium at 37o
4. Rinse the cells with DPBS.
C
waterbath prior to use.
5. Add 8 ml of DPBS first and then 2 ml of trypsin/EDTA solution into flask (in the case of
T-75 flask); gently rock the flask to make sure cells are covered by trypsin/EDTA
solution; incubate the flask at 37o
C incubator for 1 to 2 minutes or until cells are
completely rounded up (monitored with inverted microscope). During incubation, prepare
人前脂肪细胞--内脏a 50 ml conical centrifuge tube with 5 ml of fetal bovine serum (FBS, cat. no. 0500);
transfer trypsin/EDTA solution from the flask to the 50 ml centrifuge tube (a few percent
of cells may detached); continue incubate the flask at 37o
Note: Use ScienCell Research Laboratories’ trypsin/EDTA solution that is optimized to
minimize the killing of the cells by over trypsinization.
C for 1 or 2 minutes more (no
solution in the flask at this moment); at the end of trypsinization, one hand hold one side
of flask and the other hand gently tap the other side of the flask to detach cells from
attachment; check the flask under inverted microscope to make sure all cells are
detached, add 5 ml of trypsin neutralization solution to the flask and transfer detached
cells to the 50 ml centrifuge tube; add another 5 ml of TNS to harvest the residue cells
and transfer it to the 50 ml centrifuge tube. Examine the flask under inverted microscope
to make sure the cell harvesting is successful by looking at the number of cells left
behind. There should be less than 5%.
6. Centrifuge the 50 ml centrifuge tube (harvested cell suspension) at 1000 rpm (Beckman
Coulter Allegra 6R centrifuge or similar) for 5 min; re-suspend cells in growth medium.
7. Count cells and plate cells in a new, poly-L-lysine coated flask with cell density as
recommended.
Caution: Handling human derived products is potentially biohaza dous. 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 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).
Instruction for Preadipocyte Differentiation
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37o
and return them to culture as quickly as possible with minimal handling!
C waterbath
Set up of Undifferentiated Expansion of Human Preadipocytes:
4. Primary Human Preadipocytes (HPAs) should be expanded with PAM (cat # 7211) 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.
5. 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.
6. Change the medium every other day thereafter, until the culture is ready for subculture.
Induction of Adipocyte Differentiation:
8. Plate the preadipocyte suspension in PAM at a density of 10,000 cells/cm2
9. Incubate the cells at 37°C in a 5% CO
in the coated
flask or plate.
2
Note: Cells should reach 100% confluence before initiating adipocyte induction.
humidified incubator for 1-2 days.
10. When the cells are 100% confluent, carefully replace the PAM with Preadipocyte
Differentiation Medium (PADM, Cat # 7221). This medium change counts as
differentiation day 1.
11. Replace the medium with fresh PADM every 2-3 days.
12. The process of differentiation to mature adipocytes is complete after 5-12 days. Mature
adipocytes can be fixed and stained with Oil Red O Solution. Lipid droplets can be
observed after 3 days.
13. Mature adipocytes can be maintained in Adipocyte Medium (AdM, Cat. #7201) up to 6
days.
Oil Red O Staining Protocol:
0.3g Oil Red O in 100 ml isopropanol .This solution is stable for up to 1 year
Stock Oil Red O solution
The working solution
1. 3 parts of stock Oil Red O solution and 2 parts of distilled water. (Note: Let it sit at room
temperature for 10 min.
2. Filter the working solution completely through the filter funnel.
3. This solution is stable only up to 2 hours. Make it freshly every time you use it.

1. Remove media; rinse cells 2X with PBS.
Procedure
2. Fix the cells by covering 10% formaldehyde.
3. Let plates/flasks sit at least for 15 min (or overnight) at room temperature.
4. Make the working solution as described above.
5. Remove fixative solution (10% formaldehyde); gently rinse tissue culture vessels with
H2
6. Remove the water; add Oil Red O filtered working solution slowly along the side of
culture vessels. Ensure even spreading throughout the wells/flasks.
O.
7. Sit > 10 min (1 hour or longer) at room temperature.
8. Rinse with tap water until the water runs clear.
9. View the plates on a phase contrast microscope. Lipids will appear red.
Human Preadipocytes-visceral (HPA-v, Cat. # 7210) were observed under a phase contrast
microscope.
A. The cells were cultivated in Preadipocyte Medium (PAM, Cat # 7211) for 5 days (Control).
There were no lipid droplets (10X).
B. The cells were cultivated in Preadipocyte Differentiation Medium (PADM, Cat # 7221) for
5 days. Lipid droplets were detected under microscope (20X).
人前脂肪细胞--内脏A B
Caution: Handling human derived products is potentially bioharzadous. Although each cell strain tests negative for
HIV, HBV and HCV DNA, diagnostic tests are not necessarily 100% accurate, ther fore, proper precautions must
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).