Name: Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer (Video) | JoVE Journal
Uploaded: 2025-01-17T13:20:00.000+00:00
Duration: 7 min 55 s
Description: Learn flow cytometry-based isolation of tumor-infiltrating lymphocytes for adoptive cell therapy in pancreatic cancer using a mouse model. This method enables evaluation of TIL therapeutic efficacy and optimization for clinical applications.

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5 minutes

This research was funded by The National Natural Science Foundation of China (NSFC) grants, grant number 82272767 and 82072691 to Y.M. The authors thank their team members for their discussion and cooperation.

The mouse colony is housed in an AAALAC International-accredited facility, adhering to all relevant USDA, HHS, and NIH regulations and standards. The Tianjin Medical University Cancer Institute and Hospital Animal Care and Use Committee reviewed and approved all animal procedures, including the rationale for strain selection, experimental group assignments, and the statistical justification for animal numbers and randomization.
1. Tumor Induction
<ol>
	<li>Use the KPC-Luc cell line11 to induce orthotopic pancreatic tumors in mice.</li>
	<li>Prepare a cell suspension by trypsinizing the KPC-Luc cells, washing them twice with PBS, and resuspending them at a concentration of 1 x 106 cells/mL in PBS with a 30% basement membrane matrix (BMM).</li>
	<li>Anesthetize the mouse with isoflurane at an induction dose of 4% and then switch to a maintenance dose of 2%. Provide thermal support using a heating pad and monitor as necessary. Apply ophthalmic ointment to both eyes to prevent dryness.</li>
	<li>Disinfect the surgical area multiple times in a circular motion using both an iodine-based scrub and alcohol. Administer a subcutaneous injection of carprofen 5 mg/kg as analgesia prior to the surgery. Using a sterile scalpel blade, perform a small left abdominal incision (0.5-1 cm) under the left rib of the mouse to expose the pancreas.</li>
	<li>Use forceps to hold the mouse spleen and expose the mouse pancreas, mix cells and inject them into the pancreas of the mouse with 50,000 cells per mouse. Inject 50 &#181;L of the cell suspension directly into the pancreas using a 29 G needle.</li>
	<li>Return the mouse pancreas and sequentially suture the peritoneum and skin with sutures.</li>
	<li>Allow the mouse to recover on a warm pad until fully awake. Administer 5 mg/kg of carprofen subcutaneously as analgesia after surgery daily for 3 days.</li>
	<li>Perform live imaging 7 days post-injection to monitor tumor growth. 
	NOTE: Treatments may be administered at this step.
	<ol>
		<li>Intraperitoneally inject the mouse with D-Luciferin potassium salt (15 mg/mL in PBS, 60 &#181;L per mouse).</li>
		<li>Following isoflurane anesthesia, position the mouse in the in vivo imaging system for bioluminescence detection. After imaging, remove the mouse from the system and allow it to recover from anesthesia naturally.</li>
	</ol>
	</li>
</ol>
2. Harvest tumors
<ol>
	<li>Euthanize the donor mice using a CO2 chamber. Sterilize the whole body with 75% ethanol.</li>
	<li>Place the mouse in a sterile hood. Use sterile scissors and forceps to open the abdominal cavity.</li>
	<li>Remove the tumors and place them in PBS on ice. Measure the tumor size with a&#160;vernier caliper and calculate the tumor volume using the formula (&#960; x length &#215; width &#215; width)/6.</li>
</ol>
3. Tumor digestion
<ol>
	<li>Prepare the digestion solution by making 0.125 mg/L&#160;collagenase, 0.125 mg/L&#160;Dispase II, and 10 &#956;g/mL&#160;DNase I in RPMI 1640 as 1x concentration. Add 1-2 mL of digestion solution per well in a 12-well plate.</li>
	<li>Mince the tumors into small pieces using sterile scissors. Digest the tumor pieces at 37&#176;C with shaking at 70 rpm for 30 minutes.</li>
</ol>
4. Cell collection
<ol>
	<li>Terminate the digestion by adding 2-4&#160;mL of 10% fetal bovine serum. Transfer all cells to a collection tube.</li>
	<li>Wash the wells with PBS twice to collect any remaining cells. Filter the cell suspension through a 40 &#181;m or 70 &#181;m cell strainer.</li>
	<li>Collect the cells by centrifugation at 400 x g for 5 min at 4 &#176;C. Resuspend the cells in 5% BSA for flow cytometry staining.</li>
</ol>
5. Flow cytometry preparation
<ol>
	<li>Prepare spleen or peripheral blood mononuclear cells (PBMCs) from non-tumor-bearing mice as controls. 
	NOTE: Optional: PBMCs from tumor-bearing mice can also be used as controls.</li>
	<li>Stain cells with Fc blockers (&#60;1 &#956;g per million cells) and flow antibodies (CD3-APC, CD45-APC/Fire750 at a 1:100 dilution).</li>
	<li>Incubate on ice in the dark for 30 min. Wash twice with cold PBS and resuspend the cells in 1% BSA in PBS.</li>
</ol>
6. Cell sorting
<ol>
	<li>Sort CD45+CD3+ cells using a flow cytometer. Gate live cells based on FSC-A and SSC-A (P1). Then, gate single cells from P1 based on FSC-A and FSC-H (P2).</li>
	<li>Finally, sort CD45+CD3+ from P2 using CD45 and CD3 gating and collect them into the collecting tubes. Ensure proper resuspension and maintenance of cell viability during sorting.</li>
</ol>
7. Cell preparation for transfer
<ol>
	<li>Centrifuge sorted cells at 400 x&#160;g for 5 min at 4 &#176;C. Resuspend the cells in mouse serum at a final concentration of 1 x 106 cells/mL. Keep the cells on ice. 
	NOTE: Ex vivo culture or activation of the sorted cells may be performed.</li>
</ol>
8. Adoptive transfer
<ol>
	<li>Perform tumor induction for the recipient mouse 7 days before the transfer day.</li>
	<li>Inject 2 x 105 cells per mouse intraperitoneally into recipient mice using 29 G x 1/2&#34; insulin syringe. Administer IL-2 (50 IU per mouse) intraperitoneally for three consecutive days.</li>
</ol>
9. Tumor monitoring
<ol>
	<li>Perform live imaging on day 4 and day 7 post-transfer to assess tumor growth, with additional live imaging weekly with the same procedure described in step 1.8.</li>
	<li>Follow up on the survival of the recipient mice. At the end of the experiment, euthanize the mouse as described above. Use ophthalmic scissors and forceps to remove the tumors, and then place them in PBS on ice for a quick measurement of tumor size and may continue the subsequent experiments as needed.</li>
</ol>

Adoptive Therapy Using Tumor-Infiltrating Lymphocytes in Pancreatic Cancer Models

<ol><li>Ma, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combination+of+PD-1+inhibitor+and+OX40+agonist+induces+tumor+rejection+and+immune+memory+in+mouse+models+of+pancreatic+cancer%5BTitle%5D)+AND+%22Gastroenterology%22%5BJournal%5D)">Combination of PD-1 inhibitor and OX40 agonist induces tumor rejection and immune memory in mouse models of pancreatic cancer.</a> Gastroenterology. 159 (1), 306-319.e12 (2020).</li>
<li>Pitt, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Targeting+the+tumor+microenvironment%3A+Removing+obstruction+to+anticancer+immune+responses+and+immunotherapy%5BTitle%5D)+AND+%22Ann+Oncol%22%5BJournal%5D)">Targeting the tumor microenvironment: Removing obstruction to anticancer immune responses and immunotherapy.</a> Ann Oncol. 27 (8), 1482-1492 (2016).</li>
<li>Zhang, Y., Zhang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+history+and+advances+in+cancer+immunotherapy%3A+Understanding+the+characteristics+of+tumor-infiltrating+immune+cells+and+their+therapeutic+implications%5BTitle%5D)+AND+%22Cell+Mol+Immunol%22%5BJournal%5D)">The history and advances in cancer immunotherapy: Understanding the characteristics of tumor-infiltrating immune cells and their therapeutic implications.</a> Cell Mol Immunol. 17 (8), 807-821 (2020).</li>
<li>Tay, C., Tanaka, A., Sakaguchi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Tumor-infiltrating+regulatory+t+cells+as+targets+of+cancer+immunotherapy%5BTitle%5D)+AND+%22Cancer+Cell%22%5BJournal%5D)">Tumor-infiltrating regulatory t cells as targets of cancer immunotherapy.</a> Cancer Cell. 41 (3), 450-465 (2023).</li>
<li>Betof Warner, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Expert+consensus+guidelines+on+management+and+best+practices+for+tumor-infiltrating+lymphocyte+cell+therapy%5BTitle%5D)+AND+%22J+Immunother+Cancer%22%5BJournal%5D)">Expert consensus guidelines on management and best practices for tumor-infiltrating lymphocyte cell therapy.</a> J Immunother Cancer. 2 (2), e008735(2024).</li>
<li>Rosenberg, S. A., Spiess, P., Lafreniere, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+approach+to+the+adoptive+immunotherapy+of+cancer+with+tumor-infiltrating+lymphocytes%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">A new approach to the adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes.</a> Science. 233 (4770), 1318-1321 (1986).</li>
<li>Leidner, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neoantigen+t-cell+receptor+gene+therapy+in+pancreatic+cancer%5BTitle%5D)+AND+%22N+Engl+J+Med%22%5BJournal%5D)">Neoantigen t-cell receptor gene therapy in pancreatic cancer.</a> N Engl J Med. 386 (22), 2112-2119 (2022).</li>
<li>Lowery, F. J., et al. Molecular signatures of antitumor neoantigen-reactive t cells from metastatic human cancers. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aLowery%2C+FJ+%22Science+%28New+York%2C+N.Y.%29%22">Science (New York, N.Y.)</a>. 375 (6583), 877-884 (2022).</li>
<li>Rosenberg, S. A., Parkhurst, M. R., Robbins, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adoptive+cell+transfer+immunotherapy+for+patients+with+solid+epithelial+cancers%5BTitle%5D)+AND+%22Cancer+Cell%22%5BJournal%5D)">Adoptive cell transfer immunotherapy for patients with solid epithelial cancers.</a> Cancer Cell. 41 (4), 646-648 (2023).</li>
<li>Yossef, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Phenotypic+signatures+of+circulating+neoantigen-reactive+cd8%2B+t+cells+in+patients+with+metastatic+cancers%5BTitle%5D)+AND+%22Cancer+Cell%22%5BJournal%5D)">Phenotypic signatures of circulating neoantigen-reactive cd8+ t cells in patients with metastatic cancers.</a> Cancer Cell. 41 (12), 2154-2165.e5 (2023).</li>
<li>Ma, Y., Hwang, R. F., Logsdon, C. D., Ullrich, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dynamic+mast+cell-stromal+cell+interactions+promote+growth+of+pancreatic+cancer%5BTitle%5D)+AND+%22Cancer+Res%22%5BJournal%5D)">Dynamic mast cell-stromal cell interactions promote growth of pancreatic cancer.</a> Cancer Res. 73 (13), 3927-3937 (2013).</li>
<li>Mahadevan, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Type+I+conventional+dendritic+cells+facilitate+immunotherapy+in+pancreatic+cancer%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Type I conventional dendritic cells facilitate immunotherapy in pancreatic cancer.</a> Science. 384 (6703), eadh4567(2024).</li>
<li>Mullard, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((FDA+approves+first+tumour-infiltrating+lymphocyte+%28til%29+therapy%2C+bolstering+hopes+for+cell+therapies+in+solid+cancers%5BTitle%5D)+AND+%22Nat+Rev+Drug+Discov%22%5BJournal%5D)">FDA approves first tumour-infiltrating lymphocyte (til) therapy, bolstering hopes for cell therapies in solid cancers.</a> Nat Rev Drug Discov. 23 (4), 238(2024).</li>
<li>Morotti, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((PGE2+inhibits+til+expansion+by+disrupting+IL-2+signalling+and+mitochondrial+function%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">PGE2 inhibits til expansion by disrupting IL-2 signalling and mitochondrial function.</a> Nature. 629 (8011), 426-434 (2024).</li>
<li>Chupp, D. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+humanized+mouse+that+mounts+mature+class-switched%2C+hypermutated+and+neutralizing+antibody+responses%5BTitle%5D)+AND+%22Nat+Immunol%22%5BJournal%5D)">A humanized mouse that mounts mature class-switched, hypermutated and neutralizing antibody responses.</a> Nat Immunol. 25 (8), 1489-1506 (2024).</li>
<li>Stress response in tumor-infiltrating T cells is linked to immunotherapy resistance. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=%22Nat+Med%22">Nat Med</a>. 29 (6), 1336-1337 (2023).</li>
<li>Wang, A. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Glioblastoma-infiltrating+CD8%2B+T+cells+are+predominantly+a+clonally+expanded+GZMK%2B+effector+population%5BTitle%5D)+AND+%22Cancer+Discov%22%5BJournal%5D)">Glioblastoma-infiltrating CD8+ T cells are predominantly a clonally expanded GZMK+ effector population.</a> Cancer Discov. 14 (6), 1106-1131 (2024).</li>
<li>Chen, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+transformable+supramolecular+bispecific+cell+engager+for+augmenting+natural+killer+and+t+cell-based+cancer+immunotherapy%5BTitle%5D)+AND+%22Adv+Mater%22%5BJournal%5D)">A transformable supramolecular bispecific cell engager for augmenting natural killer and t cell-based cancer immunotherapy.</a> Adv Mater. 36 (3), e2306736(2024).</li>
</ol>

The authors have declared that they have no conflicts of interest. While using ChatGPT and Claude-3 for language editing, the authors carefully reviewed and validated the words and sentences generated by ChatGPT or Claude-3 before including them in the article.

Potential immunotherapy for pancreatic cancer may enhance the immune system to target and eliminate tumor cells, involving adoptive immunotherapy, immune checkpoint inhibitors, and tumor vaccines12. TIL therapy extracts and expands lymphocytes from tumors, which are then reinfused into patients. Compared to other adoptive cell therapies, TIL therapy has diverse TCR clonality, superior tumor homing ability, and low off-target toxicity, making it advantageous for treating solid tumors<sup class="xre...

Pancreatic cancer immunotherapy is in its nascent stages, with ongoing preclinical exploration and validation primarily in mouse models1. Current treatments for pancreatic cancer include surgery, chemotherapy, radiotherapy, and targeted therapies. Unfortunately, these methods often fail to completely eradicate tumor lesions, leading to recurrence and progression. Traditional treatments focus on tumor cells but often overlook the tumor microenvironment (TME), which includes both tumor cells and associated stroma composed of tumor-infiltrating lymphocytes (TILs), fibroblasts, and extracellular matrix2. TILs are immune cells within the TME, including cytotoxic T cells, helper T cells, regulatory T cells, B cells, NK cells, macrophages, and myeloid-derived suppressor cells3. These cells participate in immune responses that influence tumor growth and therapeutic outcomes4.
Adoptive cell therapy (ACT) involves collecting a patient&#39;s immune cells, modifying and expanding them in vitro, and then reintroducing them to target and kill tumor cells. TIL therapy, a type of ACT, is being researched for treating various cancers, including melanoma, lung cancer, and cervical cancer. This process involves isolating lymphocytes from the tumor, culturing them with IL-2 in vitro, and reinfusing them into the patient, often after lymphodepletion with chemotherapy or radiotherapy5.
Since Rosenberg&#39;s 1986 study demonstrated the efficacy of TILs in mice6, adoptive cell transfer immunotherapy has become a research hotspot and has shown promise in cancer treatment7,8,9,10. We aim to extract TILs from mice, sort them for specific T cells via flow cytometry, and validate their tumor-killing effects through adoptive transfer.
In this protocol, we describe a method to utilize TILs from mice through flow cytometry for adoptive cell transfer. The goal is to verify the specific cytotoxicity of TILs against tumors in a syngeneic pancreatic cancer mouse model, providing insights into the development of adoptive cell therapies for pancreatic cancer. This method includes implanting live or irradiated mouse pancreatic cancer cells in fluorescence-labeled reporter mice to initiate immune cell influx, isolating lymphocytes from primary tumors via flow cytometry sorting, or activating and expanding tumor-reactive T cells ex vivo, and adoptively transferring these activated T cells into tumor-bearing mice. Subsequent administration of interleukin-2 and bioluminescent tumor imaging allows for longitudinal monitoring of orthotopic tumor growth and response to therapy, particularly evaluating tumor-specific cytotoxic effects. This approach recapitulates the logistics involved in developing adoptive cell transfer therapies for pancreatic cancer patients. The results demonstrate enhanced antitumor efficacy of adoptively transferred tumor-reactive T cells compared to irrelevant lymphocyte controls. This versatile methodology enables the in vivo study of adoptive immunotherapy in pancreatic cancer, as well as the optimization of cell processing parameters and combination treatment regimens.

<table><tbody><tr><td>Bioluminescent imaging system</td><td>PerkinElmer</td><td>IVIS Spectrum</td><td>For monitoring tumor growth</td></tr><tr><td>Bovine Serum Albumin V</td><td>Solarbio</td><td>A8020</td><td>For cell resuspension</td></tr><tr><td>Cell strainer (40 &amp;mu;m or 70 &amp;mu;m)</td><td>Jet</td><td>CSS013040</td><td>For filtering cell suspensions</td></tr><tr><td>Centrifuge</td><td>Eppendorf</td><td>5810R</td><td>For cell collection and preparation</td></tr><tr><td>CO2 chamber</td><td>Tianjin Liliang</td><td>N/A</td><td>For euthanizing mice</td></tr><tr><td>Collagenase</td><td>Sigma-Aldrich</td><td>C5138</td><td>For digestion solution</td></tr><tr><td>CD3-APC antibody</td><td>Biolegend</td><td>100236</td><td>For flow cytometry staining</td></tr><tr><td>CD45-APC/Fire750 antibody</td><td>Biolegend</td><td>100154</td><td>For flow cytometry staining</td></tr><tr><td>C57BL/6 albino mice</td><td>Jackson Laboratory</td><td>58</td><td>Donor and recipient mice</td></tr><tr><td>Dispase II</td><td>Gibco</td><td>17105-041</td><td>For digestion solution</td></tr><tr><td>DNase I</td><td>SparkJade</td><td>AC1711</td><td>For digestion solution</td></tr><tr><td>D-Luciferin potassium salt</td><td>Beyotime</td><td>ST198-10g</td><td>For live imaging</td></tr><tr><td>Ethanol (75%)</td><td></td><td>N/A</td><td>For sterilization</td></tr><tr><td>Fc blockers</td><td>BD Biosciences</td><td>553142</td><td>For flow cytometry staining</td></tr><tr><td>Fetal Bovine Serum</td><td>Gibco</td><td>A5669701</td><td>For cell culture</td></tr><tr><td>Flow cytometer</td><td>BD Biosciences</td><td>FACSAria III</td><td>For cell sorting</td></tr><tr><td>IL-2 (Interleukin-2)</td><td>PeproTech</td><td>200-02</td><td>For administration post-transfer</td></tr><tr><td>Isoflurane</td><td>Shandong Ante</td><td>N/A</td><td>For narcotism</td></tr><tr><td>KPC-Luc cell line</td><td>PI lab generated</td><td>N/A</td><td>For inducing orthotopic pancreatic tumors</td></tr><tr><td>Matrigel Matrix</td><td>Corning</td><td>356234</td><td>For orthotopic implantation</td></tr><tr><td>PBS (Phosphate-Buffered Saline)</td><td>Servicebio</td><td>G4207-500ML</td><td>For washing and resuspending cells</td></tr><tr><td>RPMI 1640 medium</td><td>Gibco</td><td>11875093</td><td>For preparing digestion solution</td></tr><tr><td>Sterile hood</td><td>ESCO</td><td>N/A</td><td>For sterile tumor extraction and processing</td></tr><tr><td>Sterile scissors and forceps</td><td></td><td>N/A</td><td>For tumor extraction</td></tr><tr><td>Suture (4-0, PGA absorbable suture)</td><td>Jinhuan Medical</td><td>R413</td><td>For wound closure</td></tr><tr><td>Syringe</td><td>Jiangxi Fenglin</td><td>1 mL 0.45 &amp;times; 16RWLB</td><td>For injection</td></tr><tr><td>Tissue culture plate 12 well</td><td>Jet</td><td>TCP001012</td><td>For tumor digestion</td></tr></tbody></table>

flow cytometry-based isolation and therapeutic evaluation of tumor-infiltrating lymphocytes in a mouse model of pancreatic cancer

Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer

Pancreatic cancer is an aggressive malignancy with a dismal prognosis and limited therapeutic options. Adoptive cell therapy, which involves isolating and activating a patient&#39;s own immune cells, such as tumor-infiltrating lymphocytes (TILs), before re-infusing them, represents a promising experimental approach. However, techniques for adoptive cell transfer in preclinical pancreatic cancer models are not well established. Here, we describe a detailed protocol for adoptive cell therapy using TILs from a syngeneic pancreatic cancer mouse model. The procedure involves implanting live or irradiated mouse pancreatic cancer cells in fluorescence-labeled reporter mice to initiate immune cell influx, then isolating lymphocytes from primary tumors via flow cytometry sorting and/or activating and expanding tumor-reactive T cells ex vivo, and adoptively transferring these activated T cells intraperitoneally into tumor-bearing mice, followed by interleukin-2 administration.&#160;Bioluminescent tumor imaging allows for longitudinal monitoring of orthotopic tumor growth and response to therapy, especially evaluating the tumor-specific cytotoxic effects. This approach recapitulates the logistics involved in developing adoptive cell transfer therapies for pancreatic cancer patients. The results demonstrate enhanced antitumor efficacy of adoptively transferred tumor-reactive T cells compared to irrelevant lymphocyte controls. This versatile methodology enables the in vivo study of adoptive immunotherapy in pancreatic cancer as well as the optimization of cell processing parameters and combination treatment regimens.

The recipient mice are divided into seven groups to thoroughly evaluate the efficacy of the TIL transfer: (a) Saline control (blank control): Mice receiving saline injections to serve as a baseline. (b) IL-2 only (blank control): Mice receiving IL-2 injections to account for any effects of the cytokine alone. (c) The same single treatment as to the treated donor mice (positive control): Mice receiving the same treatment as the donor mice to compare direct treatment vs. TIL transfer. (d) T...

We describe a method to utilize tumor-infiltrating lymphocytes (TILs) from mice through flow cytometry for adoptive cell transfer. This protocol aims to verify the specific cytotoxicity of TILs against tumors in a syngeneic pancreatic cancer mouse model, providing insights into the development of adoptive cell therapies for pancreatic cancer.

Pancreatic cancer is an aggressive malignancy with a dismal prognosis and limited therapeutic options. Adoptive cell therapy, which involves isolating and ...

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Learn flow cytometry-based isolation of tumor-infiltrating lymphocytes for adoptive cell therapy in pancreatic cancer using a mouse model. This method enables evaluation of TIL therapeutic efficacy and optimization for clinical applications.

Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer (Video) | JoVE Journal

Flow Cytometry-Based Isolation and Therapeutic Evaluation of Tumor-Infiltrating Lymphocytes in a Mouse Model of Pancreatic Cancer

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