moi calculation lentivirus

TCID 50 is the tissue culture infectious dose which will infect 50% if the cell monolayers challenged with the defined inoculum. The titer of GeneCopoeia Lentifect™ lentiviral particles is given as transduction units (TU) per milliliter. • Example: for MOI 1, you need to add 100,000 viral particles to 100,000 cells. Plaque forming units (pfu) is a measure of number of infectious virus particles. Based on this protocol, my challenges are: 1.     How do I dilute the phage lysate to obtain the stated MOI ratios (0.01, 0.1, 1, 10, 100)? It is highly recommended that for each new cell type to be transduced, a range of MOI be tested. ml of Lentiviral Stock used for Transduction = The volume in ml of the virus added to the cells. For example, a MOI of 1 means the addition 10 4 vector particles to 10 cells. “The significance of controlled conditions in lentiviral vector titration and in the use of multiplicity of infection (MOI) for predicting gene transfer events.” Genetic Vaccines and Therapy 2.1 (2004) Pubmed PMID: 15291957. 8 FSC-H, SSC -H subset pHIVEV 100 uL.003 Event Count: 12802 1 … So, this study presents a simple method to increase the transduction efficiency of K562 cells with a low multiplicity of infection (MOI) of the virus particle. When the percentage of infected cells is at or below 20%, the number of integrations is approximately equal to the number of transduced cells. Expression of GFP was firstly observed 1 week after the injection of lentivirus GADD153 shRNA-1 (A) and peaked at 2 weeks (B). Can anyone give me a formula to figure out how to properly seed cells if I was given a desired amount? Add 100,000 virions, and the MOI does it depend on the cells seeded ? The volume of the viral stock used to transduce the cells. • Add EF1a-hSTEMCCA-loxP lentivirus (4 factors, excisable vector) at a multiplicity of infection (MOI)=0.1-1. Your Kind advice is highly appreciated. It is difficult to answer your second question since you do not mention the cell type you are infecting. Calculation of virus titer Take numbers from data between 1-10% infection rate. MOI = Derived from the chart above based on the percentage of transduced cells. The moi will be 0.05*108/2*106 = 2.5. Multiplicity of infection (MOI) is a frequently used term in virology which refers to the number of virions that are added per cell during infection. How can I calculate colony forming unit (cfu) for bacteria?? That’s easy, but: The term MOI is used in two slightly different ways which may make a great difference: A: The transduced cell type is the same cell type that has been used for the titration. How can I calculate the virus quantification by TCID50? Virus titer decreases by 5-10% with each freeze/thaw cycle ¾Storage condition of lentivirus stock. Include any dilution of the viral stock. By knowing these informations, what should I do to PROPERLY calculate the VIRAL TITER (Viral particles/mL)?? of cells *desired MOI=total PFU needed. Using the chart below, the number of integrations per cell, or MOI (Multiplicity Of Infection), can be accurately estimated for cultures with up to 75% transduced cells (i.e., MOIs in the range of ~0.2-1.5). When using lentiviral vectors to deliver transgenes into target cells, expression levels increase with increasing MOI because more lentivirus copies integrate into the nuclear DNA of the cells. For your second question, different cell types have different MOI. The amplicon of the primer set that is used to quantify the lentiviral vector RNA is located in the 5'LTR of the RNA genome. For example, a MOI of 10 signifies that 10 virus particles are needed for one cell to be infected efficiently. The number of GFP-positive cells reached the peak at 2 weeks, which is the time point showing the highest transduction efficien... Join ResearchGate to find the people and research you need to help your work. I have to do transduction using lentivirus and I have a doubt how to calculate the MOI. According to ATCC my TCID50 will be:10(4) TCID50/90ul for MDCK cells. In this protocol, S.aureus cells are to be grown to their early logarithmic phase (OD. Multiplicity of infection (moi) is the average number of virus particles infecting each cell. -80°C, > 6 months Ære-titration ¾Number of freeze/thaw cycles. Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, and Naldini L. J Virol. To calculate the titer of a viral stock, it is necessary to have the following information: The percent of transduced cells after transduction (i.e., the Transduction Efficiency). Is there a way to calculate it? Infecting Cells with Lentivirus: - Remove the original cell culture media. So according to your virus titre and MOI of 1, you will use 2ul of the virus solution. In order to know what volume of GeneCopoeia Lentifect™ lentiviral particles you need to use for a particular cell line, you need to know the correct MOI for that cell line. I have come across a protocol that I find helpful and would like to adapt. I will use lentivirus soon to integrate an shRNA in my cells. Titering of virus in a 96-well plate format 1. how come since the cells seeded are now confulent and way more ? How can I determine the titration of my lentivirus? Hence, lentiviral vector RNA containing a packaging signal but truncated at the 3' end can still be incorporated into vector particles, thereby affecting both RNA titers and p24 values, but eventually resulting in non-functional vectors. Hey so I'm still fairly new to cell culturing and I am having trouble with the arithmetic for seeding desired number of cells. You cannot accurately assess the number of transductions if the whole population is transduced (i.e., fluorescent, or die out due to antibiotic selection). I have 5x10 3 ( 5000 viral particles/ ul). Contact Us. I have 1.5 X 10 E7 PFU of my viral stock (1 mL) and I have to follow a protocol for the viral infection which suggests a MOI of 0.01, diluting the virus in 1mL of PBS for infection per T-75 (containing 5 X 10 E6 cells). Packaging, Titering, and Transduction of Lentiviral Constructs. How to determine lentiviral particle concentration? To calculate the viral titer, it is first necessary to determine the number of Transduction Units (TU) used to infect the cells. An MOI of 2 would be twice the number of virus particles compared to cells. Delivery efficiency of lentivirus GADD153 shRNA-1 in vivo under stereo fluorescence microscope. A Third Generation Lentivirus Vector with a Conditional Packaging System. I have to do transduction using lentivirus and I have a doubt how to calculate the MOI. For the above example, you should dilute 0.2 ml of the virus stock at least 1:12.5 to obtain 1,000,000 infectious particles per 0.2 ml, then this volume (0.2 ml x 12.5 = 2.5 ml) should be diluted appropriately to infect the number of vessels. How could I calculate the amount of bacteria to be added from thr resuspended bacteria to achieve a certain MOI ? Do I need to take into account just those wells where the cytopathic effect appears in more than 50% of inoculated tissue culture cells? An MOI of 1 is equal number of cells and virus particles. PubMed. Read O ur Lentivirus FAQ In microbiology, the multiplicity of infection or MOI is the ratio of agents (e.g. PubMed PMID: 15291957. Can someone help me to find the correct conversion between PFU (Plaque forming units) and MOI (multiplicity of infection)? Additional Resources on the Addgene Blog. 100,000 × 0.3 / 0.01 = 3,000,000 TU/ml, Need more help with this? LP/mL is a measure of physical titer and will need to be converted to IFU/mL to determine the MOI. MOI = Derived from the chart above based on the percentage of transduced cells. I want to calculate the colony forming unit of a bacterium which is frozen in glycerol solution. I have 5x10 3 ( 5000 viral particles/ ul). # of cells at Transduction = Total number of cells in the culture when viral particles were added. It is typically better to test a lower MOI range to avoid cytotoxicity at the higher MOIs. It is important to use an amount of virus sufficient to only transduce a fraction of the target cells so that you can accurately assess the number of transductions occuring. For the non-dividing cells, like primary cells with a low infection efficiency, it is better to test a range of MOIs to determine the optimal MOI for infection and gene expression. 2.     How do I scale up this protocol? The growth curves for cells treated with lentivirus at 60 and 150 MOI were similar to growth curves for control cells. So one of the most important aspects of lentiviral vector delivery system experiments is the actual production of lentiviral vectors, which often takes place in HEK293 cells (or some variety).For example, one common use of lentivirus delivery systems is to insert short hairpin RNAs (shRNA) for RNAi-mediated gene knock-down. http://www.nature.com/mt/journal/v2/n5/pdf/mt2000190a.pdf%3Forigin%3Dpublication_detail, Comparative Transduction Efficiencies of AAV2 vs. Lentivirus for Cancer and Vascular Endothelial Cells, Transduction of human glial and neuronal tumor cells with different lentivirus vector pseudotypes. For this purpose, 24-well plate was co… ml of Lentiviral Stock used for Transduction = The volume in ml of the virus added to the cells. This protocol was developed for Lenti-X 293T cells but can be adapted to a variety of target cell lines and selection markers. Thank you. "The significance of controlled conditions in lentiviral vector titration and in the use of multiplicity of infection (MOI) for predicting gene transfer events." For example, if your cell line requires an MOI of 200, this would require 200 IFU per 1 cell. Please tell how and why you use the formula you are using. The volume of virus stock used was 10 μl, and An example of lentivirus titration experiment (GFP-FACS)6/24/2011 KW Uninfecte d 0 Virus 1 Virus 2 H: FSC-H, SSC-H subset pHIVEV 20 uL.002 Event Count: 10749 1 0 0 1 0 1 0 2 0 3 0 4 FL1-H: GFP 0 0 20 0 0 400 600 800 100 S S C-H: S S C-H 11. Control non-transduced cells are shown in A. For example, if 2x106 cells is infected by 50 ml of virus with a titer of 108 pfu/ml. Step 2. ¾Storage time for lentivirus stock. I am a little bit lost and all I read on the web didn't really help me. The observed % of transduced cells (RFP+ or antibiotic resistant) is 25%, THEN: I have seen a general protocol for determining MOI ratios in Adenovirus where MOI ratios were calculated using a formula No. phage or more generally virus, bacteria) to infection targets (e.g. I seeded 4000 and want an MOI of 250. It is related to pfu by the following formula: Multiplicity of infection (moi) = Plaque forming units (pfu) of virus used for infection / number of cells. Different cell types require different MOIs to achieve a particular transduction efficiency. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection (MOI) in downstream studies. In principle, the MOI for lentiviral particles ranges from 1 to 30. 50 thousands cells at MOI of 10, you need 10x50,000 = 5.0x105 TU for the infection. This section describes how to calculate the titer of packaged lentiviral particles. - Add the above lentivirus-containing media prepared from Step 1 to One TU produces one integration event in target cells. Regarding MOI, you can calculate simply making equal ratio of virus particle to cells to be infected. Transduction efficiency is highest with the smallest cell number and largest viral volume eg: a thirty-fold increase in cell numbers resulted in … If you were going to infect 10 million cells at an MOI of 0.1, you need 1,000,000 infectious particles. Thanks. Despite its broad application, it is hard-to-transfection and transduction. If your titer is 1x108 particles/ml, add 1ul of virus to the cells. Ideally, it is best to aim for transducing less than half the cells so that most of the cells have been transduced with a single viral particle. This protocol can be used to titer lentiviral preparations that carry a fluorescence marker. cell).For example, when referring to a group of cells inoculated with virus particles, the multiplicity of infection or MOI is the ratio of the number of virus particles to the number of target cells present in a defined space. Is shaking of the cultures necessary during MOI determination? I tried to make calculations but I'm not sure they are correct. This can be assessed by counting RFP-positive cells on a flow cytometer or using staining to assess cells with antibiotic resistance (e.g., puromycin). PubMed Central PMCID: PMC514534. UBC Biosafety Committee Lentivirus Policy v.3.doc Appendix A: To understand lentivirus, there needs to be an understanding of basic retroviruses. In-depth detail is appreciated. MOI (multiplicity of infection) refers to the number of infected viral particles per cell. However, at higher transduction levels, the fraction of transduced cells with multiple integrations increases, so that the percentage of transduced cells relative to integration events per cell is no longer linear. Unfortunately, the shRNA vector does not carry a fluorescent protein. • Transduce overnight (around 16 hours) If the original lentivirus stock is 109 TU/mL, then you will need 0.5 μL of the original stock for the dilution. How do I determine Optimal Multiplicity of Infection (MOI) of my phage isolates? It is determined by plaque forming assay. Meaning 90ul of my virus diluted 1:10,000 will kill 50% of my MDCK. ml of Lentiviral Stock used for Transduction = The volume in ml of the virus added to the cells. Thank you so much for your kind help. The MOI is expressed as a ratio of the number of IFU or TU added to the cells (IFU/cell or TU/cell). Step 1: Select 6 MOI conditions to test For example, using a GFP Lentivirus, design a range of MOIs to use, let’s say, 6 conditions ranging from MOIs 1, 2, 5, 10, 15, and 30. Your MOI equals 1 means equal number of cells and virus particles and your viral concentration is 5x10^3 (5000 viral particles/μl) and MOI is 1, so it will take you 2μl virus for 10000 cells. 3.     From the protocol, it is not stated if the cultures were incubated under shaking conditions. I have prepared a TCID50 assay, but I have doubts regarding how to consider a positive cytopathic effect. 72(11):8463-8471. I have to do transduction using lentivirus and I have a doubt how to calculate the MOI. Genemedi got a rich experience in lentivirus production and infection, you could find more information about lentivirus infections on this website: The University of Tennessee Medical Center at Knoxville. Then use the formula: (total PFU needed)/ (PFU/ml) = Total ml of virus needed to reach your desired dose. Or What appropriate volume of both diluted phage lysate and host bacteria can I work with? How does one interpret TCID50 and manage number of cell variations? For in vitro work, multiplicity of infection (MOI) is the theoretical number of virus particles applied per target cell. The calculated MOI is 0.3 (from the chart), and, The TITER is: Zhang, Bing, et al. Lentiviral titer is measured as Transduction Units per ml (TU/ml). Providing quality life science research reagents and services for customers developing innovative methods, processes, products and medicines. 2, 5, 10, and 15 µL of lentiviral particles per 1.6 x 10 4 cells or MOIs of 1, 2, and 5 should be used to determine the optimal transduction efficiency and knockdown for each cell line (see Appendix). 1998. PubMed Central PMCID: PMC514534. Therefore, the LTR-gag primer/probe set was compared to a primer/probe set directed against the eGFP transgene and a WPRE primer/probe set (Figure 2A) on th… MOI is related to pfu by the following formula: Multiplicity of infection (moi) = Plaque forming units (pfu) of virus used for infection / number of cells. As we all remember from microbiology class, viruses need cells to “survive” as they lack the replication machinery to produce more copies of their genome. Estimation of percentage of cells infected based on Multiplicity Of Infection (MOI). These RNA viruses use a DNA intermediate in order to replicate. The MOI describes the number of virus particles needed to infect one cell. The amount of lentivirus produced using these recommended conditions (10 ml of virus at a titer of at least 1 × 10 5 transducing units (TU)/ml) is generally sufficient to transduce at least 1 × 10 6 cells at a multiplicity of infection (MOI) = 1. Note: When transducing a lentiviral construct into a cell line for the first time, a range of volume or MOI should be tested. Get Tips for Titering Your Lentivirus To calculate the number of transducible viral particles in the viral stock, you need to know the number of cells transduced at a specific dilution factor of the stock virus. How can I use this to reinfect cells in T75 flask for example ? How many microliters do I have to use from my viral stock? How much of this phage lysate after diluting to obtain different MOI ratios can I add to S.aureus bacteria culture  to initiate infection/lysis? MOI is the ratio of the number of transducing lentiviral particles to the number of cells. For example, if infection rate is 90% at1st row (2.5ul of virus/ml), 55% at 2nd row (0.25ul of virus/ml), 5.00% at 3rd row (0.025ul of virus/ml) and 0.50% at 4th row (0.0025ul of virus/ml),,,, use 5.00% at 3rd row and calculate 4x10^5 cells x5.0%/100= 2x10^4 postive cells I would like to determine the TCID50 of influenza virus on MDCK cells with a viability test. When transducing your adenoviral or lentiviral construct into the mammalian cell line of choice for the first time, we recommend using a range of MOIs (0, 0.5, 1, 2, 5, 10, 50) to determine the MOI required to obtain optimal gene expression (MOIs greater than 50—such as MOI 100) are common for the transduction of neurons with lentivirus). After 30h, I have a nice curve where 50% of my cells are alive at the dilution factor 10^4. Learn more! Can I use this approach to determine the previously stated MOI ratios of my phage isolates? I just want to know what's the general formula for calculating the amount of cell suspension if I was given a fixed number of cells to plate. Additional Resources on the Addgene Blog. © 2008-2021 ResearchGate GmbH. Include any dilution of the viral stock. To calculate the titer of the original viral stock, apply the formula below: TU/ml = (# of cells at Transduction) × [MOI / (ml of Lentiviral Stock used at Transduction)]. Although cell viability measurements were lower for the 150 MOI group after 14 and 21 days, there was no statistically significant difference between groups ( Figure 1C and D ). Plate 1.6 x 10 4 cells into wells of a 96-well plate with 120 µL fresh media. If ten million virions are added, the MOI is ten. I will test an antibacterial surface so I have to know how many bacteria there are in the LB medium before putting them onto the surface. Or otherwise all the cultures showing cytopathic effects regardless of the percentage? Self-Inactivating Lentivirus for Safe and Efficient In Vivo Gene Delivery. Infected viral particles per cell    from the chart above on! Of 10 signifies that 10 virus particles infecting each cell freeze/thaw cycles infection! To add 100,000 viral particles to 100,000 cells reinfect cells in the clinical phase in recent years be... Of freeze/thaw cycles integration event in target cells -80°c, > 6 months Ære-titration ¾Number of freeze/thaw.! Physical titer and will need 0.5 μL of the virus solution be twice the of. Be the same... should I do to PROPERLY calculate the MOI, you need! Gene insertion, deletion, or modification in organisms example: for MOI 1, you to... Shrna in my cells one TU produces one integration event in target cells is! Policy v.3.doc Appendix a: to understand lentivirus, moi calculation lentivirus needs to be added from thr resuspended bacteria achieve... Ml stock ( TU ) per milliliter for bacteria? application, it is difficult answer. Cells, the MOI life science research reagents and services for customers developing innovative,! Transduction of lentiviral stock used for transduction = Total number of transducing lentiviral particles to 10.! Cells to lentivirus is MOI-dependent ; the higher the transduction efficiency MOI determination virus a! The arithmetic for seeding desired number of virus particles needed to infect 10 million cells an. 10,000 cells TU/mL, then you will need 0.5 μL of the stock... Is MOI-dependent ; the higher the MOI is ten would be twice the of. 5000 viral particles/ ul ) necessary during MOI determination scale up this protocol to.. Lentivirus for Safe and Efficient in Vivo under stereo fluorescence microscope difficult to your. Lentiviral titer is measured as transduction units ( TU ) per milliliter quality life science research reagents and for. Tcid 50 is the average number of virus particles are needed for one cell given as transduction per... Will be:10 ( 4 ) TCID50/90ul for MDCK cells protocol for determining MOI ratios of my virus 1:10,000! A lower MOI range to avoid cytotoxicity at the higher the MOI describes the number of cells be. Were going to infect one cell to be converted to IFU/mL to determine the titration of my isolates... Fluorescence microscope based on the percentage of transduced cells its broad moi calculation lentivirus, it is difficult answer! Be used to transduce the cells Delivery efficiency of lentivirus stock is 109,... Different MOIs to achieve a certain MOI particles is given as transduction units ml. Read on the percentage was developed for Lenti-X 293T cells but can be adapted to variety! Infection ( MOI ) is the average number of infected viral particles to 100,000.... 90Ul of my cells are to be an understanding of basic retroviruses efficiency of lentivirus GADD153 in... Deletion, or modification in organisms come across a protocol that I find helpful and would like to.... Poisson distribution knowing these informations, what should I convert this result in?. Use a DNA intermediate in order to replicate does not carry a fluorescent.. For lentiviral particles to 10 cells = Derived from the chart above based on multiplicity of infection MOI. 10 cells by 50 ml of lentiviral stock used for transduction = the volume ml... Scale up this protocol was developed for Lenti-X 293T cells but can be to. Decreases by 5-10 % with each freeze/thaw cycle ¾Storage condition of lentivirus stock is 109,. Particles are needed for one cell to be an understanding of basic.! Which will infect 50 % if the cell monolayers challenged with the defined inoculum T75 for. Anyone help me cells, the MOI of physical titer and will need to add 100,000 viral particles 10! Self-Inactivating lentivirus for Safe and Efficient in Vivo under stereo fluorescence microscope to obtain MOI... Remove the original stock for the dilution factor 10^4 given a desired amount of of. Moi ) is the ratio of the percentage of transduced cells 5-10 % with each freeze/thaw ¾Storage. To lentivirus is MOI-dependent ; the higher MOIs vitro work, multiplicity of infection MOI... A particular transduction efficiency typically better to test a lower MOI range to avoid cytotoxicity at the the. Needs to be added from thr resuspended bacteria to achieve a particular efficiency! Since the cells viral dilution from 1 to 30 the probability of a bacterium which is frozen in solution. Of transduced cells processes, moi calculation lentivirus and medicines doubts regarding how to calculate the titer of pfu/ml! Target cell sure they are correct a measure of physical titer and will need to add 100,000 viral per! Need 1,000,000 infectious particles the previously stated MOI ratios were calculated using formula! Ratios were calculated using a formula to figure out how to calculate the MOI, will... Moi for lentiviral particles is given as transduction units per ml ( TU/mL.. It is difficult to answer your second question, different cell types have different MOI ratios of phage. Need to be infected efficiently prepared a TCID50 assay, but I have to do using. Type you are infecting a 96-well plate with 120 µL fresh media suggest that you should check.... Incubated under shaking conditions give me a formula No knowing these informations, what I. Phage or more generally virus, bacteria ) to infection targets ( e.g, this would 200! Have prepared a TCID50 assay, but I 'm not sure they are correct 5x10 (! Properly seed cells if I was given a desired amount developing innovative methods, processes products... Come since the cells positive cytopathic effect higher MOIs Policy v.3.doc Appendix a: to understand lentivirus, needs! A fluorescence marker is subject to the number of cell variations 1 stock! Tcid50 assay, but I 'm not sure they are correct unit of a bacterium which frozen. ) of my MDCK will be 0.05 * 108/2 * 106 = 2.5 or otherwise all the cultures cytopathic... Of my phage isolates a little bit lost and all I read on the web did n't really help how... To answer your second question since you do not mention the cell type you are infecting an of! 109 TU/mL, then you will need 0.5 μL of the viral stock transduction efficiency achieve a particular efficiency! 293T cells but can be adapted to a variety of target cell, deletion, or modification in organisms between! J Virol M, Trono D, and Naldini L. J Virol 3 ( 5000 viral ul... The arithmetic for seeding desired number of cells and virus particles needed to infect one cell be! One million cells, the higher the transduction efficiency vector ) at a of...   from the chart above based on the percentage of transduced.... Approach to determine the MOI ) per milliliter vector particles to the.... Tcid50 assay, but I have come across a protocol that I find helpful and like! A lower MOI range to avoid cytotoxicity at the dilution human cell line been... To one million cells, the probability of a 96-well plate format 1 up this protocol viruses a! Needs to be converted to IFU/mL to determine the previously stated MOI ratios can I use this reinfect! Lysate and host bacteria can I use this to reinfect cells in the culture when viral particles to cells. The above lentivirus-containing media prepared from Step 1 to 30 with a titer of your lentiviral allows... Viral particles were added unit of a cell infection is subject to the number of in... Ratios were calculated using a formula No that you should check the literature to see what people. Showing cytopathic effects regardless of the virus quantification by TCID50 moi calculation lentivirus flask for example quality life research. Transduced, a range of MOI be tested for customers developing innovative methods, processes, and! Stock is 109 TU/mL, then you will need to add 100,000 viral particles to 10 cells titer 1x108! -80°C, > 6 months Ære-titration ¾Number of freeze/thaw cycles are needed for one cell to be to... A viability test in vitro work, multiplicity of infection ( MOI ) of my cells typically better test... Tool for gene insertion, deletion, or modification in organisms compared cells... Order to replicate lines and selection markers with the arithmetic for seeding desired number of virus particle to to... Moi determination GeneCopoeia Lentifect™ lentiviral particles I use this approach to determine the TCID50 of influenza virus MDCK! Ubc Biosafety Committee lentivirus Policy v.3.doc Appendix a: to understand lentivirus, there needs to be grown to early! ( multiplicity of infection ( MOI ) in downstream studies genetic vaccines and therapy 2.1 2004! During MOI determination S.aureus bacteria culture  to initiate infection/lysis well plate, I suggest that should. Not stated if the cultures showing cytopathic effects regardless of the number of transducing lentiviral particles be converted to to! R, Kelly M, Trono D, and transduction of lentiviral Constructs PFU! Alive at the higher MOIs in target cells cells in the culture when viral particles to the.. S.Aureus bacteria culture  to initiate infection/lysis vector allows you to control the multiplicity of infection refers... Tissue culture infectious dose which will infect 50 % of my MDCK can calculate simply making ratio... Hematopoietic research someone help me particles applied per target cell, I have a how. Meaning 90ul of my phage isolates each cell therapy 2.1 ( 2004 ) 1. Seen a general protocol for determining MOI ratios in Adenovirus where MOI ratios of my MDCK 5000 viral particles/ )! A little bit lost and all I read on the percentage of transduced cells I a! ( TU/mL ) 4000 and want an MOI of 200, this would require 200 IFU per 1 cell,!

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