ISE6
CRL-3576 ™
CRL-3576 ™
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The complete medium for this cell line is L15B (see media formulation supplemental data) + 5% heat-inactivated FBS (ATCC 30-2020) + 0.1% Lipoprotein-cholesterol concentrate
ATCC Medium: L-15B
Stock A
Agitate well at a low speed until the solution becomes clear. After adding 100 µL stock A to Stock D, store in -20°C.
Stock B
Agitate well at a low speed until the solution becomes clear.
After adding 100 µL stock B to Stock D, store in -20°C.
Stock C
Agitate well at a low speed until the solution becomes clear. After adding 100 µL stock C to Stock D, store in -20°C.
Stock D
Add components listed above in order. For example, after 100 mg Ascorbic acid is dissolved, add 100 mg Glutathione. Add 5mg FeSO4.7H2O after Glutathione is dissolved. Once all of these components are dissolved well, combine this solution and 100 µL Stock A, 100 µL Stock B, 100 µL Stock C lastly to make 10 mL Stock D. After adding 1 mL Stock D into 1 L of L-15B, store at -20°C.
* Note: FeSO4.7H2O is light sensitive and rapidly oxidized by heat.
Stock E
Agitate well at a low speed until the solution becomes clear. After adding 1 mL Stock E into 1 L of L-15B, store in -20°C.
Final product L-15B
*Note: Use fresh L-glutamic acid: expired in 1 year at 4 °C
*Note: Stir slowly until all components are dissolved well without heating (< 1 hr). While stirring, protect the media from light.
Adjust pH to 7.2 by adding 10N NaOH (~0.5 mL).
Bring the volume up to 1L with Cell culture grade water.
Filter the media with 0.22 µm aPES filter unit. Aliquot 500 mL and store at 4 °C. Media expires 6 months from preparation date or the expiration date of the shortest shelf life component.
Handling Procedure for Frozen Cells
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
SAFETY PRECAUTION: ATCC highly recommends that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. Transfer the vial contents to a centrifuge tube containing 9.0 ml complete culture medium and spin at approximately 125 xg for 5 to7 minutes.
4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm2 or a 75 cm2 culture flask. It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
5. Incubate the culture at 31°C in a suitable incubator. A free gas exchange with atmospheric air is recommended if using the medium described on this product.
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
This is a patented item and order restrictions may apply. A Customer Care representative will review your order and, if necessary, will be in contact with you with further instructions. We cannot ship this item until the applicable requirements are met. We appreciate your patience during this process. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.
Munderloh UG, et al. Establishment, maintenance and description of cell lines from the tick Ixodes scapularis. J Parasitol 80: 533-543, 1994. PubMed: 8064520
Munderloh UG, Kurtti TJ. Formulation of medium for tick cell culture. Exp Appl Acarol 7(3):219-229, 1989. PubMed: 2766897
Munderloh UG, et al. Establishment of the Tick (Acari: Ixodidae)-Borne Cattle Pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae) in Tick Cell Culture. J Med Entomol (33): 656-664, 1996. PubMed: 8699463
Munderloh UG, et al. Ixodes ovatus Ehrlichia exhibits unique ultrastructural characteristics in mammalian endothelial and tick-derived cells. Ann NY Acad Sci (1166): 112-119, 2009. PubMed: 19538270
Oliver JD, et al. An Ixodes scapularis cell line with a predominantly neuron-like phenotype. Exp Appl Acarol (66): 427-442, 2015. PubMed: 25894426