AFT024 IRR細胞

數量： 大量

器官來源： 肝

細胞類型： 其他細胞類型

是否是腫瘤細胞： 0

物種來源： 小鼠

運輸方式： 凍存運輸

細胞形態： 成纖維樣

組織來源： stroma

生長狀態： 貼壁生長

年限： 14 to 14.5 day gestation embryo

ATCC Number： SCRC-1007.1?

Designations: AFT024 IRR

Depositors: ATCC

AFT024 IRR細胞Biosafety Level: 2 [Cells contain SV-40 Viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: fibroblast

Source: Organ: liver

Tissue: stroma

Cell Type: fibroblast SV40 immortalizedSV40 transformed

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Applications: It is recommended that the feeder cells be plated 24 hours before use at 5 to 6 X 106 cells/T75 in order to obtain a 100% confluent monolayer for stem cells growth.

These cells are provided for use as feeder cells to support the growth of purified mouse and human CD34+CD38- hematopoietic stem/progenitor cells.

Age: 14 to 14.5 day gestation embryo

Comments: AFT024 IRR細胞These cells are provided for use as feeder cells to support the growth of purified mouse and human CD34+CD38- hematopoietic stem/progenitor cells. They have been irradiated with 12,000 rads and will not replicate. The cells will begin to deteriorate in 2 to 3 weeks after plating. It is recommended that the feeder cells be plated 24 hours before use at 5 to 6 X 106 cells/T75 in order to obtain a 100% confluent monolayer for stem cells growth. Once the feeder cells have attached, the culture medium can be changed to accommodate the cells to be supported.

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: 2-mercaptoethanol to a final concentration of 0.05 mM; fetal bovine serum to a final concentration of 10%.

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Subculturing: Protocol: Establishing and maintaining your culture: To insure the highest level of viability, be sure to warm media to 37?C before using it on the cells. Flasks do not need to be coated before plating MEFs.

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.

Remove the vial from the water bath as soon as the contents are half way thawed (approximately 90 seconds), 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.

Transfer the vial s contents plus 5 ml of complete medium (see below for recipe) to a 15 ml centrifuge tube. Use an additional 1 ml of medium to rinse the vial and transfer the liquid to the 15 ml tube. Add 4 ml of complete medium to bring the total volume to 10 ml.

Gently mix and pellet the cells by centrifugation @ 270 xg for 5 minutes.

Discard the supernatant and resuspend the cells with 10ml fresh growth medium (warm) and transfer to one T75 flask.

Add 5 ml more fresh growth medium (warm) to flask.

AFT024 IRR細胞Incubate 37?C in a 5% CO2 in air atmosphere.

Fluid change twice a week or when pH decreases. 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 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).

Cells should be plated 24 hours before use as a feeder layer for ES cells and kept for no more than 7 days.

Medium Renewal: twice a week or when pH decreases

Preservation: Storage temperature: liquid nitrogen vapor phase

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

Recommended serum: ATCC 30-2020

Source culture: ATCC SCRC-1007

References: 29080: Moore KA, et al. Hematopoietic activity of a stromal cell transmembrane protein containing epidermal growth factor-like repeat motifs. Proc. Natl. Acad. Sci. USA 94: 4011-4016, 1997. PubMed: 9108096

39171: Miller JS, et al. Single ***** human CD34(+)/Lin-/CD38(-) progenitors give rise to natural killer cells, B-lineage cells, dendritic cells, and myeloid cells. Blood 93: 96-106, 1999. PubMed: 9864151

70208: Moore KA, et al. In vitro maintenance of highly purified, transplantable hematopoietic stem cells. Blood 89: 4337-4347, 1997. PubMed: 9192756

70209: Hachney JA, et al.AFT024 IRR細胞 A molecular profile of a hematopoietic stem cell niche. Proc. Natl. Acad. Sci. USA 99: 13061-13066, 2002. PubMed: 12226475

70210: Punzel M, et al. The myeloid-lymphoid initiating cell (ML-IC) assay assesses the fate of multipotent human progenitors in vitro. Blood 93: 3750-3756, 1999. PubMed: 10339481

70213: Thiemann FT, et al. The murine stromal cell line AFT024 acts specifically on human CD34+CD38- progenitors to maintain primitive function and immunophenotype in vitro. Exp. Hematol. 26: 612-619, 1998. PubMed: 9657136

70214: Lewis ID, et al. Umbilical cord blood cells capable of engrafting in primary, secondary, and tertiary xenogeneic hosts are preserved after ex vivo culture in a noncontact system. Blood 97: 3441-3449, 2001. PubMed: 11369635