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Mastigamoeba aflagellifera


Product category
Amoebozoa, Mastigamoebidae
Strain designation
Type strain
Isolation source
Soil of rice field at Kanagi; isolated December 1, 2006
Geographical isolation
Japan; Aomori; Goshogawara
Product format
Storage conditions
-80°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
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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.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

Handling information

Instruction for complete medium
Media: ATCC Medium 2832 may be prepared with an indicator that turns pink under low oxygen conditions. Use of rubber-seal screw caps on culture tubes helps minimize gas exchange, and filling tubes to the base of the tube neck with medium
Culture system
Handling procedure
Storage and Culture Initiation
Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen.  If liquid nitrogen storage facilities are not available, frozen ampules may be stored at or below -70°C for approximately one week.  Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C).  Storage of frozen material at this temperature will result in the death of the culture.
  1. To thaw a frozen ampule, place in a 35°C water bath, until thawed (2-3 min). Immerse the ampule just sufficiently to cover the frozen material.  Do not agitate the ampule.
  2. Immediately after thawing, aseptically transfer contents to a glass, rubber-seal screw-capped tube containing 14-15 mL ATCC Medium 2832.  Screw cap on tightly and incubate on a 15° horizontal slant at 10-30°C (20-25°C recommended for routine cultivation).
Culture maintenance
  1. Ice culture at or near peak density for 10-15 min.
  2. Vigorously invert culture 20-30 times or as necessary to sufficiently detach cells.
  3. Aseptically transfer a 0.1 and 0.25 mL aliquot to fresh tubes of ATCC medium 2832.
  4. Screw rubber-seal caps on tightly and incubate at a 15° horizontal slant at 10-30°C (20-25°C recommended for routine cultivation).
  5. Subculture when many trophozoites are observed (typically every 2-5 days).  The transfer interval will depend on the quantity of the inoculum, the incubation temperature, and the degree to which microaerophilic conditions are maintained inside the culture vessel.  This interval should be empirically determined by examining the culture on a daily basis until the growth cycle has stabilized.
Reagents for cryopreservation
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium, 8.0 mL
  1. Harvest cells from several cultures that are in the late logarithmic to early stationary phase of growth.  Place culture vessels on ice for 20-30 min.
  2. Vigorously invert tubes 20-30 times or as necessary to sufficiently detach cells, then centrifuge at 500 x g for 5 min.  Handle cultures promptly after centrifugation to avoid the amoebae reattaching to the culture tubes.  Note:  Increased yield may be obtained by using a sterile cotton swab to rub the inside surface of culture tubes both before and after centrifugation.  Use of a refrigerated centrifuge will aid in preventing reattachment of cells to culture tubes during or immediately following centrifugation.        
  3. Adjust the concentration of cells to between 5 x 105/mL - 5 x 106/mL using reduced medium (i.e., supernatant).
  4. Mix the cell preparation and the cryoprotective solution in equal portions.  Invert the tube several times to obtain a uniform cell density.
  5. Dispense 0.5 mL aliquots into 1.0 - 2.0 ml plastic sterile cryules (special plastic vials for cryopreservation).
  6. Place the vials in a controlled rate freezing unit.  Use the following cooling cycle: From room temperature cool at -10°C/min to the heat of fusion; from the heat of fusion to -40°C, cool at -1°C/min.  At -40°C plunge into liquid nitrogen.  The cooling cycle should be initiated no less than 15 and no more than 30 minutes after the addition of DMSO to the cell preparation.
  7. Store ampules in a liquid nitrogen refrigerator until needed.
  8. To establish a culture from the frozen state, place an ampule in a 35°C water bath, until thawed (2-3 min).  Immerse the vial just sufficiently to cover the frozen material.  Do not agitate the ampule.
  9. Aseptically transfer contents of thawed ampule to a glass, rubber-seal screw-capped tube containing 14-15 mL ATCC Medium 2832.
  10. Screw cap on tightly and incubate on a 15° horizontal slant at 10-30°C (20-25°C recommended for routine cultivation).  Observe the culture daily and transfer when many trophozoites are observed.


A Tonouchi
Chain of custody
ATCC <-- A Tonouchi
Type of isolate
Year of origin
Cross references
GenBank AB55091 ssu rRNA gene sequence of strain AF065-Y

Legal disclaimers

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.

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

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.


Frequently Asked Questions


Curated Citations

Uchimura Y, et al. Mastigamoeba aflagellifera sp. nov. isolated from the soil of a rice field in Japan. (in preparation for submission)

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