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Shanghai Xinyu Biotechnology Co., Ltd

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shxysw02@163.com
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15221858802,13816899465
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Room 8306, 3rd Floor, Xinjiu Plaza, 2077 Maixin Road, Songjiang District, Shanghai

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SK-N-FI human neuroblastoma cells

NegotiableUpdate on 04/24

Model

Nature of the Manufacturer: Producers

Product Category

Place of Origin

Online Consult

Overview

Cell Basic Attributes $r $n Cell Name SK-N-FI Human Neuroblastoma Cell Other Name SK-N-FI $r $n Product Code XY-H1033 $r $n Species Source Human $r $n Tissue Source Brain $r $n Growth Characteristics Adhesive Growth $r $n Cell Morphology Epithelial Cell Like $r $n Cell Specifications 1 #215; Packaging in 10 ^ 6 cells/T25 culture bottles or 1mL cryovials

Product Details

SK-N-FI human neuroblastoma cells

1、 Basic cellular properties
Cell name	SK-N-FI human neuroblastoma cells
Cell nickname	SK-N-FI
product code	XY-H1033
source organism	person
Age and gender	-
Source of organization	brain
Growth characteristics	Wall attached growth
cell morphology	Epithelial like cells
Background Introduction	-
Biosafety level	-
Cell specifications	1 × 10 ^ 6 cells/T25 culture bottle or 1mL cryovial packaging
Mycoplasma testing	none
culture medium	DMEM+ 0.1 mM Non-Essential Amino Acids (NEAA)+ 10% fetal bovine serum (FBS)
culture conditions	Gas phase:95%air+5%carbon dioxide; temperature37℃
Freezing conditions	Serum free cryopreservation solution, stored in liquid nitrogen
Doubling time	~24-30hour

2、 After cell receptionThe handling of

1)After receiving the cells, disinfect the bottle wall with 75% alcohol and place the T25 bottle at room temperature for about 1 hour. If you find that the culture bottle is damaged, there is liquid overflow, or the cells are contaminated, please take photos and contact us promptly.

2)Please confirm the cell status under a 4 or 5X microscope, and take 2-3 photos (10 ×, 20 ×) of the newly received cells, as well as one photo of the appearance of the culture bottle, for retention as a basis for the cell status upon receipt during after-sales service.

3)Adherent cells:Cells are left at room temperature for 1 hour, and their growth and adhesion are observed under a microscope. Some adherent cells may detach and form clusters due to vibration during express delivery.If the growth density of cells observed under the microscope is below 60%, the culture medium in the culture bottle can be removed (if there are unattached cells, they need to be recovered by centrifugation and resuspended into the original culture bottle), and 6-8mL of newly prepared pellet culture medium can be added and placed in the cell culture box for further cultivation. If the cell growth density reaches 70% -80% or more, the cells can be passaged. During the passage process, if cells shed due to transportation vibrations, they need to be recovered by centrifugation.

4)Method for handling cell detachment during transportation: Collect and centrifuge the detached cells, discard the supernatant, wash it with PBS, centrifuge and discard the supernatant, add 1ml of enzyme to the centrifuge tube for digestion for about 20 seconds, and thenpillcompleteAfter the digestion of the culture medium is terminated, centrifuge and spread again, and the remaining adherent cells are passaged according to the normal adherent cell passaging method.

5)Note: The transport medium (infusion medium) cannot be used to culture cells anymore. Please use a newly prepared cell culture condition according to the instructionspillcompleteCultivate cells using culture medium. After receiving the cells, it is recommended to subculture them for the first time in a T25 culture bottle at a ratio of 1:2.

IIIfineCell culture operation

1) Resuscitate cells:The following cell culture cryopreservation treatment is for reference only. The specific operating steps are mainly based on the product manual provided with the goods

Will contain1 mLThe freezing tube for cell suspension 37Quickly shake and thaw in a ℃ water bath, add4 mLMix the culture medium evenly. in1000 rpmCentrifuge under conditions3 minDiscard the supernatant and add1-2 mLBlow the culture medium evenly. Then add all cell suspensions to a culture bottle containing an appropriate amount of culture medium and culture overnight (or add the cell suspensions)6 cmIn the dish, add about4 mLpillcompleteCulture medium, overnight cultivation). On the third day, change the fluid and check the cell density.

2) Cell passage:If the cell density reaches 80% -90%, subculture can be carried out.

a)Discard the culture supernatant and use one that does not contain calcium or magnesium ionsPBSMoisturizing cells1-2Next time.

b)plus1 mLDigestive fluid（0.25%Trypsin-0.02%EDTA）In the culture bottle, allow the digestive fluid to infiltrate all cells, and place the culture bottle in37Digestion in a ℃ incubator1 -3min(depending on the digestion of the cells), then observe the digestion of the cells under a microscope. If most of the cells become round and fall off, quickly take them back to the operating table, tap the culture bottle a few times, and then add2-3mlpillcompleteTerminate digestion of the culture medium.

c)Gently mix and place into a sterile centrifuge tube,1000 rpmcentrifugation5 minDiscard the supernatant and add more1-2 mLBlow the culture medium evenly.

d)Press the cell suspension1:2Proportionally divided into new categories8 mLPlace the culture medium in a new dish or bottle and incubate it in an incubator.

Attention points for passaging adherent cells:

lmustPBSWash it once.

lObserve the cells several times to control the time. Do not stop digestion and blow them down before the cells have basically shed. This way, the cells will More prone to differentiation and death.

lDon't keep blowing and beating cells

3) Cell cryopreservation:When the cell growth is in good condition, cell cryopreservation can be performed. belowTTaking 25 bottles as an example;

a)Collect cells and cell culture medium, transfer them into sterile centrifuge tubes,1000 rpmCentrifuge under conditions4 minDiscard the supernatant and usePBSClean it once and discard it completelyPBSPerform cell counting.

b)Add serum-free cell cryopreservation solution based on the number of cells to achieve a cell density of 1×10^6~1×10^7/mLGently mix well and freeze each tube for storage1mLCell suspension, pay attention to labeling the cryovials properly.

c)Place the cryovial in-80℃ refrigerator,24 hThen transfer to liquid nitrogen storage. Record the location of the freezer for future retrieval.

4Cultivation precautions

1)If there is a problem with the cell, it should be resent

a)Various problems encountered during cell transportation, such as cell loss, bottle damage, severe leakage of culture medium, etc., require resending;

b)Regarding the issue of cell contamination, please provide us with authentic experimental results within 3 days of receiving the product, verify them, and resend them;

c)After allowing the cells shipped at room temperature to stand for 2 hours, and after 2 days of recovery from dry ice freezing, the vast majority of the cells did not survive. (True and clear photos of the cell status should be provided), they should be resent;

d)After 2 days of revival of cells stored in dry ice for shipment or 2 hours of standing at room temperature for shipment without opening, if contamination occurs, resend;

e)Regarding cell viability issues, please provide us with real experimental results within 7 days of receiving the product, identify cell viability using the trypan blue staining method, and provide daily cell photos for verification before resending;

f)Please take photos on the day the cells are received, as well as on the 2nd and 3rd day. If you do not inform us within 3 days, it will be considered that the product is qualified. If there is a problem within 4-7 days, please provide photos taken 3 days before receiving the cells, photos of the cell problem, and detailed steps of the cell related operations, and communicate with the technical personnel. If the technical personnel determine that it is our responsibility, it will be resent.

2)The situation where cells have problems and are not resent

a)Customer operation causing cell contamination, not resend;

b)Serious operational errors by the customer resulting in poor cell status will not be resent;

c)If the cell state is not good due to our recommended cell culture system, it will not recur;

d)The cell status is not good, and no clear and authentic photos of the cell status from the first 3 days of culture have been provided. It will not be resent;

e)If other treatments during cell culture cause problems with the cells, they will not be reproduced;

f)If the time for receiving cells and discovering problems and communicating with customer service personnel is more than 7 days, it will not be resent;

g)Depending on the specific situation

5、 Precautions

1. All animal cells are considered to have potential biological hazards and must be operated in a secondary biosafety platform. Please pay attention to protection, and all waste liquids and containers that have come into contact with these cells must be sterilized before disposal.

2. It is recommended to always use protective gloves, clothing, and a face mask when reviving frozen cells. Attention: The cryotube immersed in liquid nitrogen will leak and gradually fill with liquid nitrogen. When thawing, the conversion of liquid nitrogen into gas phase may cause the container to explode or the lid to be blown off with dangerous force, resulting in flying debris and causing personal injury.

3. This cell is for scientific research purposes only! The manual is for reference only. The actual delivery manual shall prevail!