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微信小章| 产品名称： | Acanthamoeba pearcei Nerad et al. |
|---|---|
| 商品货号： | TS154065 |
| Strain Designations： | 205-1-AX |
| Biosafety Level： | 2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Isolation： | axenic clone derived from the type strain, ATCC 50435, 1994 |
| Product Format： | frozen |
| Type Strain： | no |
| Medium： | ATCC® Medium 712: PYG w/ Additives |
| Growth Conditions： | Temperature: 25.0°C Duration: axenic Protocol: ATCCNO: 50436 SPEC: This strain is routinely shipped as a frozen stabilate. Thaw the ampule, aseptically transfer the material to 5.0 ml of ATCC medium 712 in a 16 x 125 mm screw-capped plastic test tube. Screw the cap on tightly and incubate on a 15 degree horizontal slant at 25C. Trophozoites should attach to the bottom surface of the test tube. Within 7-10 days the trophozoites will form the equivalent of a monolayer on the bottom of the tube. To subculture, vigorously agitate the tube and aseptically transfer a 0.1-ml aliquot to a fresh tube of medium. Incubate as above. Note: The transfer interval will depend on the quality of the medium, the size of the vessel, the volume of the medium, and the inoculum size. When cultivated at ATCC at 25C in plastic 16 x 125 mm screw-capped test tubes (with the caps screwed on tightly) containing 5 ml of medium 712, the transfer interval is every 2 weeks when the inoculum is 0.1 ml. Cells remain viable for longer periods in plastic than in glass test tubes. |
| Subcultivation： | Protocol: ATCCNO: 50436 SPEC: This strain is routinely shipped as a frozen stabilate. Thaw the ampule, aseptically transfer the material to 5.0 ml of ATCC medium 712 in a 16 x 125 mm screw-capped plastic test tube. Screw the cap on tightly and incubate on a 15 degree horizontal slant at 25C. Trophozoites should attach to the bottom surface of the test tube. Within 7-10 days the trophozoites will form the equivalent of a monolayer on the bottom of the tube. To subculture, vigorously agitate the tube and aseptically transfer a 0.1-ml aliquot to a fresh tube of medium. Incubate as above. Note: The transfer interval will depend on the quality of the medium, the size of the vessel, the volume of the medium, and the inoculum size. When cultivated at ATCC at 25C in plastic 16 x 125 mm screw-capped test tubes (with the caps screwed on tightly) containing 5 ml of medium 712, the transfer interval is every 2 weeks when the inoculum is 0.1 ml. Cells remain viable for longer periods in plastic than in glass test tubes. |
| Cryopreservation： | 1.xa0xa0 To achieve the best results set up cultures with several different inocula (e.g. 0.25 ml, 0.5 ml, 1.0 ml).xa0 Harvestxa0 cultures and pool when the culture that received the lowest inoculum is at or near peak density. 2.xa0 If the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107cysts/ml with fresh medium.xa0 If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration. 3.xa0 While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows: Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.xa0 Allow the DMSO to solidify.xa0 Add the required volume of refrigerated medium.xa0 Dissolve the DMSO by inverting the tube several times.xa0 xa0xa0xa0xa0xa0 *NOTE: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium. 4.xa0 Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/ml and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 min. 5.xa0xa0 Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6.xa0xa0 Place the vials in a controlled rate freezing unit.xa0 From room temperature cool at -1°C/min to -40°C.xa0 If the freezing unit can compensate for the heat of fusion, maintain rate atxa0xa0xa0xa0xa0xa0xa0 -1°C/min through the heat of fusion.xa0 At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. xa0Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.xa0 (The cooling rate in this apparatus is approximately xa0xa0xa0xa0xa0 -1°C/min.) xa0 7. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer. 8.xa0xa0 To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2-3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial. 9.xa0xa0 Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 ml of fresh ATCC medium 712 in a T-25 tissue culture flask or plastic 16 x 125 mm screw-capped test tube.xa0 Incubate at 25°C. |
| Name of Depositor： | TK Sawyer |
| Year of Origin： | 1994 |