Stem Cell Biology - Science topic

I'm just wondering if I need to change the media of my cells that are culturing in 5% O2 (hypoxic condition) in a hypoxic chamber aka a hypoxic glove box? Or can I just quickly do this in a normal incubator and return the cells to the hypoxic incubator?

It sounds like a lot of work if we need to get a hypoxic glove box so I'm just wondering if anyone has any experience in this who could give me some insight?

Thank you

I am wondering if there are any commercially available fluorescent stains for live imaging of astrocytes derived from hiPSCs. We are co-culturing neurons and astrocytes and would like to stain the live cells to determine the change in their populations over time.

I am planning to expand my business and hope to learn some industry information from you in your area. Or share some resources and explore potential cooperation opportunities.

#Medicine #Biotechnology #Doctor#Research

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Hello! I currently have differentiated iPSCs I wish to clean up. I use mTESR with matrigel coating.

ReLeSR selectively dissociates healthy stem cells, however even on quick release (1 minute), all the cells immediately dissociate, and I am unable to isolate healthy stem cells.

I think that it might be due to me passaging very regularly (once every three days), and the cells have been adapted and now dissociate easily.

I would also like to attribute that to the reason why my iPSCs are differentiated. I supplement them with ROCK inhibitor (5uM) every time I passage. Given that they are passaged once every three days, that means they are in ROCK inhibitor once every two days... could this be the reason my iPSCs are so differentiated, as I did not give them sufficient time to recover from ROCK inhibitor?

However, the reason why I passage so regularly is because I know need to passage once I see differentiated cells... Do you suggest I completely omit ROCK inhibitor during passaging, and would it help the maintenance of my iPSCs? For individuals with experience using ROCK inhibitor during passaging, do you feel that frequent passaging causes spontaneous differentiation of iPSCs even when you remove ROCK inhibitor the next day (before 24 hours)?

I would really appreciate any advice that would help me out of this predicament, thank you so much!

I am trying to grow Kasumi-1 cell line. I use RPMI 1640 and 20 % FBS but cells never seem to be growing. They keep on shrinking in size and eventually die off.

The prevailing theory is that cancerous tumors reflect a form of cellular natural selection, where random mutations confer evolutionary advantages to tumor cells. These survival benefits accumulate over time, eventually empowering tumor cells to outcompete healthy cells and defeat the immune system.

Given this theory, one expects a correlation between cell division rate and cancer rate -- tissues with the highest number of cell divisions should show the highest incidence of cancer (environmental and hereditary factors notwithstanding).

However, few papers seem to verify this assumption, or even document cell division rates and lifetime cell divisions among different tissues.

These are the two best papers so far:

However, the same senior author leads both papers, and the papers seem to omit the most common cancer types, breast and prostrate. Furthermore, the methodology seems fragile as they conducted an analysis across different studies that may not have employed uniform methods.

Could anyone recommend more robust or authoritative papers on the subject of cell division rates?

Within a new project I want to stain collected tissue from rats with alizarin red. Since I only found protocols for in-vitro applications or when tissue mentioned then only parafin slides - I would be glad if you could share with me your expertise.. The collected tissue will be mounted and frozen and cut into cryosections (this cannot be changed due to other reasons) and should be stained afterwards with Alizarin Red solution. How about the demasking,ect..?

Thank you a lot in advance..

I came across two papers (Shcheglovitov et.al., 2013, Nature; Zaslavsky et.al., 2019, Nat Neuro) where they mixed neurons from two genotypes and seeded them on a bed of rodent astrocyte. How does one use this setup as a readout for any phenotypes?

I am growing organoids in monolayer but honestly couldn't manage to make it confluent. the protocol i followed made use of collagen. they first look "like confluent" with about 70-80% cell, but after washing or changing media, they just turn from 40 to 10% remaining cells.

i thought there could be a problem with collagen? what is the best collagen concentration and how much collagen should be used to coat per well? Thank you.

I've been working with some bone marrow derived MSCs I got from Texas A&M and I've wondered what other's experience has been with bone marrow derived MSCs (or MSCs from adipose tissue, hUCB etc. doesn't matter). In my experience they tend to be slow growers (plated at ~2000 cells/cm^2 it'll take ~10 days to reach 80% confluency) and senesce quickly (I get like 7-10 passages out of them). I change the media weekly and don't use a pH indicator. I've been using stemlife MSC medium for undifferentiated expansion. This media is interesting to me because I assume its probably pretty basic but it contains what they call "life-factors" which is proprietary of course.

Has anyone on here created their own media or found a particular method of culturing to be the most robust and productive when working with bone marrow derived MSC's?

Hi, I am using a commercial line of NSCs from X Cell Science. I would like to know if anyone has stained it for PAX6 marker? If so, was it positive? Having said that, are NSCs supposed to express PAX6? What if they don't?

I routinely functionally assess the iPSC derived neurons using calcium imaging with Fluo-4/AM (490nm/510nm). However, have not been successful at using those same neuronal dishes for carrying out ICC. The reason that I think could be are as follows:

Any suggestions or references or direction to a protocol would be of a great help!

Thanks!

I wish to look at gliosis in a dish, basically the neurons start becoming apoptotic and glial cells come to the rescue. But if I have a patient neurons, how do I mimic this event? Any idea what can be used a control?

I'd like to make some Wnt/R-Sponding media to lower the costs of organoid maintenance

Many thanks

I read a study (link below) where they cultured cells and re-suspended them in a buffer before injection. I am curious what buffers are safe for this? PBS? aMEM? What do you think?

I am looking for different methods of mammalian stem cells freezing and thawing. Both in industrial and laboratory scale. Would greatly appreciate your input.

Hey guys, I have try several transfection reagent (Xfect, lipofectamine 3000, PolyJet) for transfecting mouse ES cells. But the transfection efficiency is quite low. The best I can get is 50% (using polyJet) which was transfecting a plasmid with ~10kb. But I could just get this percentage once and got much lower efficiency in the other trials (using the same plasmid). I am not sure if the reagent in our hands function properly or not because some of them are quite old and the our freezer was broken down previously which may make the reagent degrade. 

Could anyone share some experience in transfecting mouse ES cells? For example, which reagent you used? How big is your plasmid? And what is the transfection efficiency you normal get? Thank you!

Update: Cells are growing fine in our lab now!

I'm the only person in my lab starting on iPSc culture (we received a vial from another lab and all I have is some the medium they use for culture) and I'd like some tips and help. I have human iPS cells, and I culture in mTeSR1 on matrigel coated plates (so feeder free), replacing media daily. I try to remove what I think is differentiation by marking on the plate while looking through a microcscope. One cell line shows nice colonies like I saw on suppliers webpages (Like Stemcell or Invitrogen), the other I'm not so sure... 

I've attached some pictures, can someone tell me if this is what it's supposed to look like? And can give someone tips on how to passage? I've passaged one with gently scraping, like i saw mentioned in literature and tried picking the colonies with a p1000 tip (but didn't know if I actually picked up a colony or not, so I tried scraping). Maybe i just need a bit more patience :) 

While awesome to start things up, it's frustrating having no one to ask if things are supposed to be this way :)

Which concentration of Matrigel you use for iPSC culture? I was using 500 ul in 15ml and then, I have added 2 ml in 6 cm dish. However, my colonies did not attach.

Now, I am using 1.5ml of matrigel in 12 ml and so far, nothing.

:)

Hi all. I came across a technical question from one of my friends, that whether a given culture of hESCs can be inserted with three different reporter constructs using CRISPR/Cas9 method, which means a triple knock-in line hESC. I could not find any relevant answer from the literature that I came across and thought researchgate would be an appropriate platform. Hence, I request all to give your inputs in this regard. Thank you.

Differential gene/protein expression profiles on same cell line always shows varied results, why does this happen?

Do they acquire neuron-like morphologies in your cultures as well? What signalling mechanisms are taking place when they're seeded at lower cell density?

Hi

I used two cell lines hESC cutured in E8 medium feeder-free condition.When I convert into naive state use protocal by 3iL or NHSM media,I find the colonies shows continuous apoptosis and cannot passage even though forming a little dense colonies as picture.First I think the high concentration of every inhibitor harms to cells in consideration of individual cell line,but when I reduce the dose,it does not work.

Does anyone have similar experience or can give me advices?

Many thanks.

Hi everybody. I've bought human CD34+ derived iPSC line from Life Technologies. In the initial recovery as soon as I got the cells, it worked great. However when I cryopreserved the cells with E8 medium with 10% DMSO and thaw them, I found massive cell death.

Here are the details:

Day 1. Thaw cells as fast as possible. Culture them with E8 medium + ROCK inhibitor.

Day 2. Most of the cells are viable. Change the medium, with ROCK inhibitor.

Day 3. Start to see modest cell death. There're still some decent looking colonies. Change the medium, with ROCK inhibitor.

Day 4. Massive cell death. Although there're still some cells left (but not quite look like colonies) adherent in the dish (vitronectin coated), I am worry that eventually all the cell will die.

So is that normal or I did something wrong with the culture? I will definitely try longer culture.

I would like to perform a live/dead staining on my bone marrow aspiration samples, as well as add some fluorescent-labeling to track certain components. However, the auto-fluorescence of the bone marrow itself is masking everything.

Also, I can't seem to find any information on the fluorescence spectrum of bone marrow or blood.

I've read a number of protocols on this and several people's comments, but try as we might our lab is struggling to make this a success; whether we are culturing to differentiate macrophages (conditioned L929 media) or dendritic cells (GM-CSF), we're left with virtually zero viable cells at the end. I suspect that the hematopoetic progenitors are not surviving the process.

After I collect the bone marrow, I wash them once with complete DMEM and then resuspend in 90% FBS + 10% DMSO.  Aliquot the cells (10^7/tube) into cryotubes and then into rate-controlled freezing container overnight and transferred to liquid N2 the next day.  For recovery, we put the frozen tube into 37 deg waterbath until *almost* thawed, transfer the cell slurry into pre-warmed complete media (~20-30 mL), centrifuge to remove DMSO, and resuspend in differentiation media. 

The only things I can think of that might still be affecting our results is 1) the concentration of cells/freezing media may be too high or too low, 2) thawing is too quick (although I thought that was the point), 3) handling is too rough; the cells need a rest period before differentiation.

But truthfully, I have no idea. By all accounts I've heard, we should at least be getting SOMETHING. When we prepare macrophages or DCs from fresh bone marrow, all is well and our yield is excellent. 

Does anybody have any thoughts? What the heck are we missing here?

Hi,

I have successfully isolated stem cells from fresh urine but I am trying to figure out if the urine sample can be refrigerated for a period of time equivalent to shipping on ice and still get cells out. I am setting up an experiment to try and isolate cells from urine that is refrigerated for 24, 48 and 72 hours but I was wondering if people have done this. I haven't found many papers online. Any suggestions are welcome.

Thanks!

Elisa

I am establishing a mo-DC protocol, and there is too many option in regards of maturation. I would like some help deciding. Since later in downstream applications I would be combining IFNg, I would like to include this cytokine in the protocol. However, I've seen that in order to truly create maDC i should activate the other canonical pathways. That is why I though of using TNFa and LPS. Before jumping and trying it out, I was wondering if anyone has experience with this type of cocktail or IFNg+TNFa.

Thanks

Good morning.

I study IPSC-derived mesenchymal stem cells (MSC).

After differentiation into MSC, cell proliferation speed was slow than normal MSC.

Cells did not proliferate after 4 passages and their morphology changed (aging phenotype?).

I already asked. But I still have not solved the problem. I do not want to add growth factors.

Can I use only the MSC Media Supplement FBS for MSC extensions?

Thank you in advance.

I am trying to culture mouse embryonic stem cells in 3D chitosan hydrogel but can't harvest cells from hydrogel in order to do some assays like cell count or cell viability,etc. is there any effective method to detach cells from hydrogel without damaging them?

Normal epithelium has a turnover. Life span of the epithelial differentiated cell is short. For example, lifespan of colonic epithelium cell is 3-4 days. Even within this short period, the differentiated epithelial cells are constantly in contact with the stem cells. Somatic stem cells generate differentiated progenies at regular basin to maintain tissue homeostasis. This turnover is regulated by feedback or feed forward regulation. In feedback regulation, stem cells take feedback messages (information) from their progenies. In particular, stem cells seem to send feedforward signals to their progenies to regulate their behavior .

Quantum entanglement is a physical phenomenon that occurs when pairs or groups of particles are generated or interact in ways such that the quantum state of each particle cannot be described independently of the others, even when the particles are separated by a large distance—instead, a quantum state must be described for the system as a whole.(Wikipedia)

If we think genes as quants  can we explain stem cell-progeny relation (feedback, feedforward)with quantum mechanics?

hi, anyone with experience on the potential toxic effects of nano-scaled materials on stem cells in vitro or/and in vivo?

The emerging evidence suggests that the drawbacks of senescence are twofold. First, one might expect, senescence causes a loss of tissue repair capacity because of cell cycle arrest in progenitor or stem cells. Second, senescent cells produce pro-inflammatory and matrix-degrading molecules (such as IL1-beta, TNF-alpha and MMP, respectively) in what is known as the senescence-associated secretory phenotype (SASP). The SASP is largely initiated by NFkB and p38MAPK signal pathways. I wonder what kind of other signal pathways are involved in SASP? JNK signal, for example, seems to be too transient to cause SASP. 

Dear colleagues,

i am planning to track down the uptake and processing of a FITC-labeled antigen (e.g. FITC-OVA) by Peyer's Patches after oral gavage. Unfortunately, most of the literature deals with antigens bound to particulated matters, like nanoparticles (chitosan, etc.). Is it possible to track the uptake of a soluble antigen by Peyer's Patches as well? I plan to isolate Peyer's Patches cells and use FACS analysis or fluorescence microscopy along with T cell markers to look for Th1 or Treg shift, respectively.

Thanks in advance!

Hello all,

I'm looking to start using an ALDH assay as a marker of stem-like characteristics in Cancer Stem Cells (CSCs). These assays are quite expensive and before I commit to purchasing one, I was wondering if anyone had first hand experience with assays such as Aldefluor or Aldered. (Or any others) Do they work well? Is there an assay anyone considers to be the best or better than the rest?

Thank you for your time.

I'm growing human primary myoblasts & tenocytes in 3D collagen type I gel, where I see huge increase of doubling time in the 3D gel, up to about 4 fold increase.

Has anyone had this issue and is this a normal behaviour for these cells in 3D collagen gel?

Thank you.

I need to send my stem cell samples to another institution. What is the time limit? One of my colleagues  said 8 h is safe for tissue samples. However, I didnot come across such an information. How about stem cells? And is there any paper that I can cite?

I am culturing gastric cancer cell with 10% FBS. From last few months i have noticed some black dots floating in the plated media and slowly the number of dots getting increased. After reviewing some article i got a paper which suggests, these black dots are achromobacter which can be eradicated by 10mg/L ciprofloxacin and piperacillin in combination. but still i have question about PSA which we normally use as disinfectant. when we add 0.1% PSA along with 10mg/L Ciprofloxacin and Piperacillin, is it good for cellular health?  

We have noticed that some patients' cells freeze and thaw easily and those patients get good clinical response to frozen cells. There is a group who do not respond well to frozen cells. In the vet world we see some bulls whose semen will not freeze. Is this the same for human SVF cells?

Dear Colleagues!

Recently I've performed TEM observation on cultured spermatogonial cells. I've found two unknown structures (attached below). I thought, they're just artifacts, but they reappeared several times during my observations. I tried to find analogical structures elsewhere, but to no avail. Does anyone knows what could it be? I'll be grateful for any advice.

I am working at the problem of mammalian development and stem cell differentiation. And wonderingly how can we define different cell types, and how many cell types in a specie. Any one know where can I find the answers?

I will add this solution to collected menstrual blood, and then isolate endometrium stem cell from blood. So this solution had better to be sterile.

Hello, some questions..mammalian cells are usually frozen between 1 x 106 cells/mL to 1 x 107 cells/mL. do you know the best concentration/cell density to freeze differentiated stem cells?I've observed that they do not like to divide as well they do after several passages. I am wondering if the amount of cells per vial could affect on it in terms of cell death or may be they just dedifferentiate. Thank you

can we stoped the aging of cells

Q1. I am planning to use an ALP kit to detect the mES cells pluripotent/ differentiation status. Which would be the better (ALP) kit for this purpose?

Q2. I generally passage mES cells every 2-3 days. But some kit says 4-5 days (crucial for ALP activity). The detection days (4-5) are very strict? If grow the cells for 5 days, the cells were not looking good., may be started differentiated (EB?).  

(I used 2i+LIF medium with supplements)

Q3. Does anybody having experience of using- Vector Blue Alkaline Phosphatase Substrate Kit III (from Vector Laboratories) for mES cells?

Thanks in advance!

I am culturing mESC RW4 cells and found that they start to differentiation after two passages.

For culturing, I used the knockout DMEM (Gibco), with 15% FBS (ES cell, Gibco), 2mM L-glutamine, 10mM Hepes, 0.1mM non-essential amino acid, 0.1mM beta-mercaptoethanol. The media is prepared freshly and used up to 1 week. Fresh LIF is added to the cell directly every time when changing media or splitting. I change the media everyday for mESC. And of course I used 0.2% gelatin for coating the plate and have MMC treated fibroblast as feeder cell for mESC.  

I attached the figure to show the morphologies of mESC at p1 (left figure) and p3 (right figure). mESC at p1 are undifferentiated with nice round shape. mESC at p3 have two types of cells mixed together. I think the round colonies are still undifferentiated. But those small round shining cells around the colonies look neither fibroblast nor mESC. To me, they are differentiated stem cell. I will use AP1 staining to confirm if they are really differentiated or not in the next step. 

I am so upset they are easily to differentiate. Due to the consideration that the FBS batch are not good, I have tried to change the FBS(optimized for ES cell) to knockout serum replacement (Gibco)  and at the same time change trypsin to accutase (Gibco). I have also asked another lab for borrowing the good FBS that have been tested by them. These changes did not help with maintaining the stem cell status.

 I am now desperate and would like to ask if anyone has experience the same situation and if any one could suggest any element or staff that I should be aware. Thanks a lot!!

I have read that (as usual with kits!) the amount used can be significantly dialled down to make the kits go a little further. Does anyone have experience with this?

Hello everybody.

I'm trying to culture and differentiate murine DPSC into neurons like cells, but the proliferation ratio is really low.

Moreover, present cells show different morphologies (no genetic analysis done yet).

Do you know any coating for the plate that would increase proliferation and differentiation into neurons like cells?

Thanks ;)

Does anyone have experience on differentiation of mouse induced pluripotent stem cells (iPSCs) into hematopoietic fate? I need an experimental design to differentiate my iPSCs into hematopoietic lineage but not a specific lineage. I need to know which culture supplements are the minimum requirements and finally which markers and functional tests should be analyzed to confirm the successful differentiation (which genes and surface markers must be analyzed by qPCR and flowcytometery?). owing to the limited grant, I appreciate the answers with the minimum compulsary markers. thank you. 

Hi, I am interested in doing pluripotency analysis of my mESCs, and want to include the earliest markers for differentiation. I do observe some form of differentiation in my colonies, but not sure as to which lineage they could belong to. I am looking for something that's probably before the neural lineage, but ahead of naive pluripotent nanog/oct4 expressing stage?

Many thanks.

AraC doses.

I use the 4034 from stemcell. Seed cell are cd34+ from hES. 

my cells doubling time around one day and i cultured them four days ago and they did not double or increase in number. they did not die neither. may be the medium is not enough? .

We are using alpha-MEM with 10% FBS and 1% penstrip. But growth is very slow.

I want to know if ILC3s specifically, and more broadly all ILCs can be reconstituted by adoptive transfer of bone marrow cells from a widl-type mouse to a lethally irradiated knockout mouse.

My aim is to isolate as much RNA as possible from a small amount of mesenteric arteries of rat (less than 100 um diameter) 4 fragments (less than 100ug) which then will be used for further cDNA experiment. Is that possible with this quantity?

Could I get any help what is the best Kit or Protocol to achieve the best results with the lowest amount of sample.

Dear all,

I'm trying to stain actin filaments with Phalloidin Tetramethylrhodamine B isothiocyanate (TRITC) on C3H10T1/2 Cl8 cells (mouse embryo fibroblast).

I'm following a well established protocol of my lab. Cells are:

PBS washed

fixed with paraformaldehyde 4% 10 minutes

PBS washed

permeabilized with 0.1% Triton 10 minutes

 PBS washed

incubated with 50 ug/ml phalloidin in PBS for 30 minutes

PBS and finally water washed

air dryied and mounted with glycerol:PBS (9:1)

In the same experiment (same cells) in some glass coverslips are clearly visible actin fibers, well stained, while in other glass coverslips nuclei seem stained and instead of clear actin fibers I can see just a "cloud" of actin around the nuclei. Furthermore, boundaries of cells seem not well attached.

I repeated the experiment several times and I always have some samples not well stained (or fixed).

Does anyone has an idea of which experimental parameter I'm ignoring?

I have the feeling that is something related to fixation, but I'm confused about the nuclear actin. I read that phallodin shouldn't bind to nuclear actin, unless cells are stressed (Nuclear actin dynamics – From form to function, 2008), but I'm ignoring which stress cou ld be implied.

 

Thanks in advance.

Proliferation rate using MTT assay 

I wish to work on mesenchymal dental pulp stem cells.. I need to know the best procedure so that the viability of stem cells is maintained when extracted from dental pulp.

I want to transduct mouse Adipose Tissue-Derived Mesenchymal Stem Cells with lentiviral that expression GFP buttransduction don't succeed, GFP expression is not detectable .

I use working concentration for polybrene is 8 to 10 μg/ ml.microgram/ml) for transduced For 12 to 14 hours.

do cells need some pre-treatment before adding antibodies?

I am working with human NSCs and trying to differenciate them, I was recomended PLL as a good coating material. Does someone have experience with PLL or PDL and could tell me something about it. Thanks!

We have a patient who developed ascites with G-CSF during mobilization and I wonder if any body has had such an observation. 

I am using gefitinib to inhibit the EGFR. I want to perform RT qPCR on EGFR target genes to make sure the drug is actually working.

i want to differentiate mesenchymal stem cell into neuron 

do you advise me to use bone morphogenic protein 9 or not .

Hi everyone. I've been working with MACS for isolation of UCB CD34+ cells for quite sometime and now I have a problem in my hands that I cannot understand or solve. Usually after the isolation, the cell viability is high (at least 80%) and somehow now,me and my colleagues are getting viabilities of 50% and lower.

I have done a systematic study where I have change a set of parameters:

- change Microbead kit

- change column lot

- change type of column (LS and MS)

- Use higher concentration of DNAse for thawing MNCs (50ug/mL)

- change MACS buffer: 2%FBS + 2mM EDTA, instead of 0.5% BSA

- Put MNCs in complete media overnight at 4ºC or at 37ºC

Still, no improvements were seen...

We use frozen MNCs in recovery media, that were obtained from Ficoll density gradient separation.

Can please anyone have an idea of how to overcome this? Did this ever happen to anyone?

Thank you for your help!

Márcia

I'm working tissue engineering and need Tilapia scale collagen powder raw material for fabrication scaffold , any body knows which company sell that?

The epithelial-cell layer that lines the intestine often needs to be repaired, a process that is initiated by intestinal stem cells, which reside in the crypts in the intestinal wall.

We are puzzled by a case where we excised part of the small bowel transplant and pathology showed extensive ulceration, with only 5% of mucosa preserved, and absence of crypts. Serosa and muscle layers are well preserved.

Would MSCs help regenerate the mucosa? Do you believe the absence of crypts is prohibitory?  

Can stem cell implants help schizoaffective disorder?  If so, what is the closest location to Richmond, Virginia?

I was wondering if anyone could provide me with some information about the cryopreservation of human bone marrow stem cells. 

If you are doing the slow-freeze method, what is the largest volume you can successfully freeze? Every article I've come across always mentions volume sizes of 1-2 mL, and are usually vague about anything larger than that. 

I've also seen some variance in optimal freezing cell concentrations. 

Is it possible to freeze a volume of cells in the 125-250 mL range? What preserving reagents do you recommend (serum-free)? What cell concentration would be recommended?

I'm performing a western blot testing for alpha-SMA. I purchased the antibody from abcam (ab5694), and was wondering if it could be incubated at room temperature; and if so, how long it should be incubated? 

My understanding is that certain antibodies can degrade at room temperature. Therefore, I wanted to see if someone had already figured out the optimal time for primary antibody incubation. 

Thanks! 

Hello.

As the title describes, we are looking for the BL21 E.coli strain to use it for a demonstration and experimentation with bacteria and protein isolation in some high school classes in Biology lesson.

Does anyone know a way to get access at those cells ? Is there any institute or university or even a person that can send in Europe and more specific in Greece some cells in a micro-tube ? 

Thank you very much.

Hello everyone,

I currently use Sendai virus to reprogram my fibroblasts and PBMC to iPSCs. As I have translational aspirations, I need to switch to other approaches. Any thoughts about episomal plasmids? or other non integrative methods? What do you do in your labs?

Thank you!