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About

The Cell Culture Facility, located in 336 Barker Hall, supports research by providing cell cultures (mammalian and insect), media, supplies and expertise on a recharge basis for more than 40 different laboratories on and off the main campus. The facility provides cell cultures for researchers to take to their labs for experiments, provides media and materials for checkout, and also allows use of laminar flow hoods, incubators and microscopes for researchers who are using cells provided by the Facility, to use on site. See our services section for more details below.

    Services

    • Expert consultation for researchers regarding cell culture needs, media, and experimental design
    • Grow and culture mammalian and insect cells, and the entire Allen Institute genome-engineered human iPSC cell collection
    • Expansion and harvesting of hybridoma lines
    • Cell Authentication by STR analysis provided in conjunction with the DNA Sequencing Facility (More Info)
    • Mycoplasma screening
    • User training in cell culture technique
    • Cryopreservation of cell lines
    • Provide tested media, serum, and cell lines
    • Provide and stock cell culture reagents including media, serum, additives, and other reagents
    • Use of 2 laminar flow hoods for mammalian or insect cell manipulation*
    • Contract work for outside companies

    *Major equipment includes five biosafety cabinets; two phase contrast microscopes, one fluorescence microscope seven CO2 incubators, one mixed gas incubator and two incubators for insect cell lines; two incubators for large scale growth of cells using roller bottles or spinner flasks; cold storage facilities consisting of cryogenic storage dewars, one ultra low freezer, four -15° C freezers, two refrigerators and a cold room for storage. The core is located in three rooms in Barker Hall with easy access for most researchers in UCB.

    Ordering and Hood Reservations

    *All new clients and undergraduates must meet with us before using our hoods to review standard operating procedures (undergraduates must be working with a graduate student). ALL undergraduates must also meet with us for AT LEAST one cell culture training session before independent use. Undergraduates may tour the facility with a graduate, however, they must still meet with us prior to independent use.

    Rates

    Hourly rate: 

    University of California Laboratories: $68/hr *

    External laboratories (academic and industry): $110/hr 

    *Most orders can be completed in 15 minutes for a total of $17 (ie 6-well plate orders, T-75 orders, 10cm dish orders etc)

    Vials: 

    University of California laboratories: $68/vial

    External laboratories (academic and industry): $110/vial *

    *Shipping and dry ice fee not included

    Available Cell Lines and Media

    List of Cell Lines (XLSX)
    List of Media (PDF)

    If you don't find the cell line you're interested in please email cellculture@berkeley.edu.

    If you order cells from ATCC, you can deposit them with the Cell Culture Facility. We can expand your cell line and store multiple vials in our Facility.

    If you want to deposit a non-ATCC cell line in our facility please refer to the Protocols page of our website and review the Mycoplasma Screening protocol. If you can provide the Facility with a fixed, mycoplasma free sample on a coverslip we will gladly accept your line.

    CRISPR Cell Line Repository

    Published CRISPR interference, activation, or knockout cell lines may be provided upon request from the Cell Culture Facility at UC Berkeley. To learn more and get up-to-date information on available lines, please contact Alison Killilea at cellculture@berkeley.edu.

    A list of available cell lines is available here:

    Allen Institute Human Induced Pluripotent Stem Cells (iPSC)

    The UC Berkeley Cell Culture Facility is excited to offer many fluorescently tagged hiPSC lines created by the Allen Institute. The Allen Cell Collection has 52 high quality-certified fluorescently tagged hiPSC lines that target 41 key cellular structures and substructures available to help your research program. You can find supplies for culturing these cells including matrigel, media, rock inhibitor at our facility.

    All of our hiPSC banks have been validated using STR, Gband analysis, and markers for stemness at our lab. They have also been screened for mycoplasma.

    Please make sure your lab has BSL2 approval on your BUA and that you have SCRO (Stem Cell Research Oversight Committee) approval before placing any orders (click here for more information).

    Additional information regarding certificate of analysis can be found here. Culturing protocols, images, and videos can be found in our protocol section. 

    Please note that an MTA is required for the distribution of these stem cell lines, which you can find here.  

    For University of California stem cell laboratories: 

    Our MTA with the Coriell Institute allows us to sell any hiPSC cell line from the Allen Institute Cell Collection to other labs in the UC System at a significant discount. Please contact us for our rates.

    Synthego Gene Editing of IPSC lines

    A partnership between the UC Berkeley Cell Culture Facility and Synthego has been established in order to provide researchers at UC Berkeley with greater access to Synthego’s iPSC Engineering services. To place an order, please go to our ordering section.

    Protocols

    Cell Culture Protocols

    Insect Cell Protocols

    Mycoplasma Screening

    MTT Assay

    UCB Core Facilities for Analyzing Your Cells

    Publications and Grants

    Publications incorporating core services should credit the UCB Cell Culture Facility which is supported by The University of California Berkeley. Our RRID is SCR_017924. A resource statement for grant applications is available below:

    The Cell Culture Facility provides mycoplasma testing, cell authentication using short tandem repeat analysis, growing of cultured mammalian and insect cells, the entire Allen Institute for Cell Science genome-engineered human iPS cell collection and cell storage services. In addition, the Facility trains users in cell culture techniques and provides sterile hood facilities for users. Major equipment includes five biosafety cabinets; two phase contrast microscopes, one fluorescence microscope seven CO2 incubators, one mixed gas incubator and two incubators for insect cell lines; 2 incubators for large scale growth of cells using roller bottles or spinner flasks; cold storage facilities consisting of cryogenic storage dewars, one ultra low freezer, four -15° C freezers and 2 refrigerators and a cold room for storage. The cell culture facility stocks cell culture reagents including media, sera, additives, and other reagents. The core is located in three rooms in Barker Hall with easy access for most researchers in UCB.

    Contact Information

    Staff:
    Core Facilities Director: Alison Killilea
    Staff Research Associate: Willie S. Hercule
    Undergraduate: Lynda Solis Chavez
    Undergraduate: Wendy Fang

    For services and questions, please contact us by email at cellculture@berkeley.edu or by phone at 510-664-4053.

    After Hours Access

    Barker is unlocked during business hours and card access is not needed M-F 7am to 5pm.

    If you need to access our facility after hours please fill out this form. You must have a Berkeley ID. Please allow up to one week for your ID to be processed. 

    Frequently Asked Questions

    Q: When is the latest I should place my order for: 

    An adherent line with a desired confluence?

    A:  9am the day before you would like to be ready. Please note we only keep a certain number of plates per cell line. For larger orders (i.e. more than 1-2x10cmTCD or more than 1x15cmTCD), try to place orders at least 3 days before your desired pick up date. Additionally, if we are not currently growing the cell line, we may need additional time to make your order. We will let you know of any delays in advance. 

    An adherent line with a desired cell count?

    A: Before 9am the same day you would like it to be ready. Please note that if the cell count is higher than the amount we have in stock or if we are not currently growing the cell line, we will let you know in advance. 

    An insect cell suspension line? (Insect lines are split M,W,F)

    A: HIGH5: before 9am on the split day you would like it to be ready. For orders greater than the equivalent of 1x1L @ 1e6, please place your order by 9am the split day prior to your pick up date (e.g. A 2L HIGH5 order with a density of 1e6/ml for pick up on a Wednesday should be placed by 9am the Monday before. A Monday pick up for the same order should be placed by 9am the Friday before). 

    SF9: before 9am on the split day you would like it to be ready. For orders greater than the equivalent of 2x1L @ 1e6, please place your order by 9am the splitting day prior to your pick up date (Assuming no larger orders are in front of your order).

    *for pick up dates that are not on a split day (e.g. Tues or Thurs), you should place orders as if they are to be picked up on the split day prior (e.g. A HIGH5 order <1L @1e6/ml for pick up Tuesday should be placed Monday by 9am. A HIGH5 order >1L @ 1e6/ml for pick up Tuesday should be placed Friday by 9am.)

    A mammalian suspension line? 

    293 GNTI (split M,W,F): before 9am the splitting day prior to your pick up date

    EXPI F: (split M, Th): before 9am the splitting day prior to your pick up date

    *Please note larger orders (293 GNTI: >100ml @ 1e6, EXPI F: >125ml @ 1e6) will take additional time. These orders should be placed at least a week in advance. But we suggest at least 2 weeks especially for orders 293 GNTI orders larger than the equivalent of 2L @ 1e6/ml)

    Q: Do I need to do anything after I place my order?

    A: If you plan to use recycled flasks for your suspension lines, please bring them at least two splitting days in advance. We do not keep black 96 well plates. If you request them for an order, you must provide your own a day in advance. 

    Q: When will my order be ready?

    A: You can expect your order to be ready on your desired pick up date. For same day orders, pick up is after 2pm. If there is a delay, we will inform you ahead of time. For mycoplasma and testing and STR validation, please allow up to a week before results. 

    Q: Will you contact me when my order is ready? 

    A: No. Once you place your order, expect that it will be ready by 2pm on the day you requested. The only exceptions are suspension lines that require us to thaw the same day. Those will be ready by 4:30pm at the latest. We will only contact you if there is a reason for delay or need to clarify a request. 

    Q: When will I receive my shipped order?

    A: Next day. We provide overnight shipping Monday-Thursday (we don't ship on Friday). As long as we have your lab's chartstring, FedEx account number, and shipping information, orders placed before 11:30am will be received the next day. If there's an issue or delay, we will let you know ahead of time.

    Q: What passage is my cell line?

    A: Cell line passages can be found in the upper left corner of your dish. It will be marked with a “P” followed by the passage number. 

    Q: What passage is my frozen cell line?

    A: You can request that information when you place your order and we will let you know when you come by to pick it up. 

    Q: How do I pick up my order?

    A: Find the pick up shelf (insect cells are in the insect cell incubator next to Freezer 1 or in small order incubator located on the counter across from the insect cell incubator on the right side of the large centrifuge. Mammalian suspension lines are in incubator 1 or 2, adherent lines are in incubator 3). Put on gloves and spray with ethanol. Open the incubator and pick up your order. If your order is on a shaker DO NOT TURN OFF THE SHAKER when picking up your order. All other mammalian cell line orders will be in incubator 3. 

    Q: Do I need to bring anything when picking up my order? 

    A: EHS requires that you bring a secondary container to place any live cells. If you are coming to pick up frozen cells, please bring dry ice. If you forget, we will direct you to the stock room on the ground floor where you can pick up dry ice. 

    Q: Do I need a code to get in? 

    A: You will need a code to get in before 9am and after 5pm. If you do not have a door code, please email cellculture@berkeley.edu

    Q: When can I access items in the freezer?

    A: Before 3pm M-F. 

    Q: Can I store my cell line with you? 

    A: Yes. However, all cell lines must be mycoplasma tested. Please follow the mycoplasma screening protocol on our website. 

    Q: What cell lines are available? 

    A: Please refer to our list of cell lines on our website under our “Available Cell Lines and Media” section. 

    Q: What media do I use for my cells? 

    A: Please refer to our list of cell lines on our website under our “Available Cell Lines” section for required media.

     

    Troubleshooting 

    Q: Why are my cells dying after a thaw?

    A: There are a number of reasons for unsuccessful thaws. 

    1. Media: Certain cell lines are very sensitive to the type of media being used. If the media isn’t exact, they could die. Please make sure your cells are being thawed in the proper media and check our xlsx file under Available Cell Lines for media conditions.

    2. They’ve been left in DMSO: DMSO is a cryopreservative that can be toxic to cells when left thawed for too long. Our thawing protocol is designed to get rid of DMSO expeditiously since cells are quite sensitive when recently thawed. If not followed correctly, cells can begin to die. Cells are frozen in 10% DMSO so adding our cells to 9 mls of media only dilutes the DMSO to 1% which can still be toxic to cells. Best practice is to pellet the cells and resuspend in fresh media.

    3. Contamination: Yeast and bacteria are common contaminants when thawing cells. Note that our protocol is also designed to help mitigate risk of contamination. 

    4. Incubator: CO2 and temperature levels are critical for fresh thaws. Please make sure that the CO2 is circulating properly, at 5% and the temperature is at 37 degrees Celsius when placing your cells in the incubator. 

    5. If you’ve checked these four things and none of them seem to be the issue, you can email us for further consultation at cellculture@berkeley.edu 

    Q: Why aren’t my cells lysing after viral transfection?

    A: Although it is rare, it does occasionally happen. As long as your cells are still producing the desired protein, the protein should still be usable.