Human Primary Neural Stem Cells

Cat. No.
T4178
Unit
5x105 cells / 1.0 ml
Price
Inquiry
Cat. No. T4178
Name Human Primary Neural Stem Cells
Organism Human (H. sapiens)
Tissue Brain
Donor History Normal tissue
Growth Properties Suspension
Cell Type Primary Cells
Unit 5x105 cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions

Thaw cells in non-tissue culture 10 cm petri dishes. The growth media for this cell line is PriGrow X Series Medium (TM4178) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Change half of the medium every 2-3 days by tilting flasks and allowing neurospheres to settle in one corner. 

TM4178 contains TM4178-D (Neurosphere Dissociating Solution) which is required for subculturing cells.  

Primary Neural Stem Cells may be differentiated into the following lineages with their respective differentiation media:

  • Neuron Lineage Differentiation Medium (TM4178-N)

  • Astrocyte Lineage Differentiation Medium (TM4178-A)

  • Olgiodendrocyte Lineage Differentiation Medium (TM4178-O)

Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.


Thawing Protocol

1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 9ml of pre-warmed, complete growth media. Centrifuge cells at 200xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in 5 ml of the recommended pre-warmed, complete growth media and dispense into a 10 cm petri dish with 10 ml of complete growth media.

5. Incubate the cells at the recommended conditions.

6. Cells should float in the complete frowth media (TM4178) and will proliferate to form floating neurospheres in 2-3 days. Detach any neurosphere that attach to the culture vessel with gentle pipetting. Do not allow neurospheres to attach to culture vessel; dislodge several times a day as neccessary. 

7. Change half of the media every 2-3 days by tilting the flask and allowing the neurospheres to settle in one corner. Remove half of the media and repalce with the same volume of fresh, complete growth media. 

8. Dissociate neurospheres when they reach 100-200 um in diameter using the Neurosphere Dissociating Solution (TM4178-D).  

Subculture Protocol

Dissociate neurospheres when they reach 100-200 um in diameter using the Neurosphere Dissociating Solution (TM4178-D).  

1. Warm the Neurosphere Dissociating Solution to room temperature. 

2. Collect neurospheres in a 15 ml conical tube and centrifuge at 800 rpm for 3-5 minutes. 

3. Carefully aspirate the supernatant and re-suspend the pellet in 3-5 ml of pre-warmed Neurosphere Dissociating Solution. Optional:  add 30-50 ul of DNase I. 

4. Incubate in a 37.0°C water bath for 2 minutes. Neutralize the dissociating solution with 10 mL of complete growth medium. 

5. Centiruge at 1200 rpm for 5 minutes. Carefully aspirate medium and resuspent neurospheres in 1-2 ml of growth medium. 

6. Gently triturate 10-15 times with pre-wetted fire polished glass pipette to break up the neurospheres. 

7. Allow any undigested spehres to settle by gravity and transfer the cell suspension to a new tube. 

8. Add 1-2 ml of the growth medium to the undigested clusters and repeat step 6 and 7. Combine the cell suspension and plate into two non-tissue culture 10 cm petri dishes.


Primary Neural Stem Cells may be differentiated into the following lineages with their respective differentiation media:

  • Neuron Lineage (TM4178-N)

  • Astrocyte Lineage (TM4178-A)

  • Olgiodendrocyte Lineage (TM4178-O)


To differentiate cells:

1. Dissociate neurospheres as described above.   

2. Dilute the neural stem cell suspension using the complete growth media (TM4178) to 100,000 cells per ml. 

3. For differentiation, seed cells at 50,000 cells per cmin culture vessels pre-coated with Poly-D-Lysine and Laminin Coating Solution (ratio of coating solution to surface area = 1ml per 5cm2).

  • Incubate culture ware with coating solution for a minimum of 1 hour. Aspirate coating solution. Wash coated surface three times for 15 minutes per wash with sterile PBS.

4. Incubate cells growth media (TM4178) overnight 37.0°C, 5% C02.

5. Change media to the Differentiation Medium of your choice the next day. 

6. Check the culture daily and change half of the differentiation medium every other day. 

7. Observe differentation into matured neruonal cells of your choice using an inverted microscope. Cells will differentiate to:

  • 7-10 days for astrocytes;

  • 2-3 weeks for neurons

  • 4 weeks for oligodendrocytes

Cryopreservation

Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

Seeding Density (cells/cm2) 36,000
Split Ratio 1:2
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T4178.
Warranty abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”.
Disclaimer

1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].

2. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

3. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

4. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.

5. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."

Application Research Use Only.
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T4178
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