上海11选5走势图

主页 > 美国ATCC > 干细胞
价格:请临时咨询
数量:
ATCCACS-1029 ATCC-BXS0115 Human [Hispanic Female] I ..
基本信息
资源编号 ATCCACS-1029
资源名称 ATCC-BXS0115 Human [Hispanic Female] I ..
种属 Homo sapiens, human Derived from Bone Marrow CD34+ cells
类型 sendai virus reprogrammed hiPSC
提供形式 frozen
致病类型 Normal
安全等级 2
培养方法
培养基 ATCC iPSCs have been adapted to feeder- and serum-free culture conditions.
The base medium for this cell line is Pluripotent Stem Cell SFM XF/FF (ATCC® No. ACS-3002) which is a ready-to-use medium for serum-free and feeder-free iPSC culture. 
传代方法 Cell culture dishes are coated with CellMatrix Basement Membrane Gel (ATCC® No. ACS-3035) to provide a surface for the attachment of iPSCs.
Coating Procedure: Thaw CellMatrix Gel on ice and swirl gently to mix. Important: CellMatrix Gel will solidify in 15 to 30 minutes above 15°C. Keep CellMatrix Gel, vials and pipette tips on ice at all times to prevent CellMatrix Gel from solidifying. If air bubbles form, they may be eliminated by centrifuging CellMatrix Gel at 300 x g for 10 minutes at 2°C to 8°C. Determine the appropriate volume per aliquot based on concentration and usage. Example: 2 mL of CellMatrix at 150 μg/mL is required to coat one 6-cm dish. To coat two 6-cm dishes, prepare as follows: Dilute CellMatrix in DMEM:F12 to a working concentration of 150 μg/mL. For instance, if the protein concentration of CellMatrix (on certificate of analysis) is 14 mg/mL, then: (4 mL) x (0.15 mg/mL)/(14 mg/mL) = 0.043 mL. Therefore, add 43 μL CellMatrix directly in 4 mL cold DMEM: F-12 Medium Cell culture dishes coated with CellMatrix Basement Membrane Gel should be incubated at 37°C for one hour. Aspirate coating solution and immediately plate the cells. It is critical that the coating does not dry out. Volumes used in this protocol are for a 75 cm2 flask. Post thaw day 1, perform a 100% medium change and remove all cells that did not attach. Perform a 100% medium change every day. Passage the cells every 4 to 5 days (80% confluent) at an appropriate split ratio (a 1:4 split ratio is recommended). If the colonies are close to, or touching each other, the culture is overgrown. Overgrowth will result in differentiation. ROCK Inhibitor Y27632 is not necessary each time the culture medium is changed. It is required when cells are recovering from thaw; it is recommended when the cells are being passaged. This protocol is designed to passage stem cell colonies cultured in a 6 cm dish, using EDTA Dissociation Reagent to detach the cell colonies. The recommended spilt ratio is 1:4. Volumes should be adjusted according to the size and number of the tissue culture vessels to be processed.  EDTA Dissociation Reagent:  500ul 0.5M EDTA 0.9g NaCl in 500ml Calcium/Magnesium free PBS Sterile filter and store at 4°C Note: Addition of ROCK inhibitor has been shown to increase the survival rate during subcultivation and thawing of human iPSCs. The use of ROCK inhibitor may cause a transient spindle-like morphology effect on the cells. However, the colony morphology will recover after subsequent media change without ROCK inhibitor. Warm an aliquot of EDTA Dissociation Reagent working solution to room temperature. Aspirate and discard the stem cell culture medium. Rinse the cells twice by adding and discarding 4 mL of D-PBS. Add 2 mL of EDTA Dissociation Reagent working solution to the dish. Incubate at 37°C for 2 to 5 minutes. Aspirate the EDTA Dissociation Reagent and gently rinse the colonies with 4 mL of DMEM: F-12 Medium, taking care not to dislodge the cells during manipulation.  Aspirate the DMEM: F12 rinse and discard. Add 2 mL of stem cell culture medium with ROCK Inhibitor Y27632 to the dish, and detach the cells by pipetting up and down 2 to 3 times with a 1 mL tip. Take care not to over-pipette the culture into a single-cell suspension as single cells will not establish colonies after seeding.  Transfer the cell aggregates to a 15 mL conical tube. Add an additional 3 mL of stem cell culture medium with ROCK Inhibitor Y27632 to the dish to collect any remaining cells. Transfer this rinse to the 15 mL conical tube containing the cell aggregates. Centrifuge the cell aggregates at 200 x g for 5 minutes. Aspirate the supernatant and discard. Add 1 mL of stem cell culture medium with ROCK inhibitor Y27632. Gently resuspend the pellet by pipetting up and down 2 to 3 times with a 1 mL tip, maintaining the small cell aggregates. Take care not to over-pipette the culture into a single-cell suspension as single cells will not establish colonies after seeding. Plate the cells as desired on feeder or feeder-free cultures. Incubate the culture at 37°C in a humidified 5% CO2/95% air incubator. Perform a 100% medium change every day. Passage the cells every 4 to 5 days (80% confluent).
生长条件 Pluripotent Stem Cell SFM XF/FF, ATCC ACS-3002 CellMatrix Basement Membrane Gel, ATCC ACS-3035
ROCK Inhibitor Y27632, ATCC ACS-3030
Stem Cell Dissociation Reagent, ATCC ACS-3010
存储条件 Liquid Nitrogen Vapor Phase (-130°C or colder);For optimal results, cryopreserve stem cell colonies when the cell cultures are 80%confluent. This protocol is designed to cryopreserve stem cell colonies cultured in a 6 cm dish. Detach stem cell colonies from the dish as described in the recommended subculturing protocol (steps 1-11). Gently tap the bottom of the tube to loosen the cell pellet. Take the Stem Cell Freezing Media from storage and swirl to mix. Keep cold. Decontaminate by dipping in or spraying with 70% alcohol. Add 2 mL of cold Stem Cell Freezing Media (ATCC ACS-3020) to the tube. Gently resuspend the pellet by pipetting up and down 2 to 3 times with a 1 mL tip, maintaining the cell aggregates. Immediately transfer 1 mL each of the cell suspension into two labeled cryovials. Freeze the cells gradually at a rate of -1°C/min until the temperature reaches -70°C to -80°C. A cryopreservation container (e.g., CoolCell® freezing container) may also be used. The cells should not be left at -80°C for more than 24 to 48 hours. Once at -80°C, frozen cryovials should be transferred to the vapor phase of liquid nitrogen for long-term storage.
详细说明
Age 24
Gender Female
Ethnicity Hispanic
Karyotype Normal karyotype, 46 XX
Derivation ATCC-BXS0115 Human Induced Pluripotent Stem Cells (iPSCs) were derived from bone marrow CD34+ cells obtained from a healthy Hispanic female donor.
Antigen Expression SSEA4, Tra-1-60 (expressed on undifferentiated hiPSCs) >85%; SSEA1 (expressed on differentiated hiPSCs) < 15%.
Cells per Vial ≥30 colonies after 5 days when seeded as directed
STR Profile Consistent with expected
Sterility Tests No growth after 21 days
Mycoplasma - None Detected
Zero Footprint Confirmation - residual Sendai virus not detected
Viral Testing Hepatitis B Negative
HPV Negative
HIV1 Negative
CMV Negative
EBV Negative
Functional Tests Pluritest - Pluripotency Score >20, Novelty Score
Year of Deposit 2013
FAQ's iPSC - ROCK inhibitorDate Updated 1/23/2014 iPSC - Karyotypic instabilityDate Updated 1/23/2014 iPSC - iPS cells following passageDate Updated 1/23/2014 iPSC - Long recovery time of iPSCDate Updated 1/23/2014
Restrictions This product may be used by investigator for research purposes subject to the Limited Use Label License and any additional third party terms. This product is subject to claims under U.S. Patent Nos. 8,058,065 and 8,048,999, pending patent applications, and foreign counterparts thereof.  In addition, this product was generated using the proprietary technology of the DNAVEC Corporation, including, without limitation, recombinant Sendai virus vectors. For information on obtaining additional rights, please contactlicensing@atcc.org
山西11选5开奖 上海11选5走势图 云南11选5走势图 58彩票开户 万家彩票开户 上海11选5 内蒙古11选5走势图 内蒙古11选5开奖 印象彩票平台 一线彩票平台