AUCTORES
Globalize your Research
Research Article | DOI: https://doi.org/10.31579/2640-1053/009
*Corresponding Author: Zhang H , Biomedical Engineering from Beijing Institute of Technology
Citation: Zhang H, Role of Biorepositories in Personalized Medicine: Isolation and Processing of Primary Tumor Cells for the initiation Tumor Cultures, J Cancer Research and Cellular Therapeutics, Doi: 10.31579/2640-1053/009
Copyright: © 2017 Zhang H. This is an open-access article distributed under the terms of The Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Received: 20 March 2017 | Accepted: 15 April 2017 | Published: 05 May 2017
Keywords: Three-dimensional tumor culture; Personalized medicine; Primary tumor cell culture; Tissue repository, Drug sensitivity testing
The introduction of three-dimensional (3D) tumor cultures has revolutionized anticancer drug research as these cultures allow for the study of drug resistance mechanisms that cannot be explored in traditional two dimensional (2D) monolayer cultures. Discoveries in the 3D tumor culture field suggest that individualized drug sensitivity testing of solid tumor specimens through the establishment and use of 3D tumor cell cultures following tissue collection will become a routine service offered by modern tissue repositories as they expand from their traditional research role to active participation in personalized medicine. Unfortunately, most information related to 3D tumor cultures comes from studies using established tumor cell lines rather than primary tumor cultures. However, accumulation of genetic aberrations in cancer cell lines occurs with increasing number of passages severely limiting their usefulness for personalized medicine. There is only very limited information available concerning technologies and standard operating procedures for the efficient and routine isolation and processing of primary tumor cells for the establishment of 3D tumor cultures from solid tumor specimens. The purpose of this work was to review experimental data from the literature that may provide relevant information concerning the isolation and processing of primary tumor cells for the establishment of 3D tumor cultures. Information reviewed here may help bio repositories in the development and standardization of technologies and standard operating procedures related to the use of 3D tumor cultures.
Accumulation of genetic aberrations in cancer cell lines occurs with increasing number of passages severely limiting their usefulness for personalized medicine [1,2]. In contrast, the critical importance of technologies related to the derivation, short term culture, and testing of primary tumor cells from solid tumors is increasingly recognized (for a recent review see 2). Short-term primary cultures of tumor cells derived from pieces of solid tumors have been used for predicting anticancer drug responses [3,4]. Observations suggest that individualized drug sensitivity testing of solid tumors through the isolation and culturing primary tumor cells in 3D may soon become routine and modern tissue repositories will need to be ready to support these activities as they expand from their traditional research role to active participation in personalized medicine [5].
Cancer cells grown in 3D cultures in a polymeric ECM closely mimic the biology of tumor development in vivo and numerous studies indicate that 3D cultures are superior to traditional 2D monolayer cultures for studies of key cellular behaviors like differentiation, proliferation, invasion and apoptosis [6-8]. Cancer cells grown in 3D culture are more resistant to chemotherapeutic agents, radiation and oncolytic virotherapy than cells in 2D culture and 3D tumor cell cultures are useful for preclinical evaluation of the cytotoxic effect of anticancer agents [6, 9-16].
Despite the significant advances in our understanding of various aspects of cancer initiation, progression, metastasis, tumor microenvironment and cancer stem cells we have achieved limited clinical success. According to the current figures published by the American Cancer Society, more than half a million people die every year due to cancer in the USA alone.
A review of the English language literature related to the isolation and processing of primary tumor cells for the establishment of 3D tumor cultures from solid tumor specimens was performed.
Only a limited number of reports are available related to the isolation and processing of primary tumor cells for the establishment of 3d tumor cultures from solid tumor specimens. However, these relate to colorectal, lung and brain tumors indicating that isolation and growth of primary tumors cells in 3D cultures is possible for a variety of solid tumors. The protocols have an initial step of mechanical mincing of tissues followed by at least some disruption of the original extracellular matrix by enzymatic dissociation. Isolated tumor cells or fragments can be grown and analyzed in a wide variety of 3D culture systems. Four prominent examples are reviewed below.
Kondo et al found that retaining cell–cell contact enables preparation and culture of spheroids composed of pure primary cancer cells from colorectal cancer specimens that retain the characteristics of the parental tumors, grow in vitro in 3D cultures and could be used for personalized diagnostic applications, including chemosensitivity assays.
To successfully maintain the cell–cell contact, they used Liberase DH (Roche Diagnostics) as a blend of digestion enzymes. They optimized the enzyme concentration and the digestion time to avoid overdigestion of colorectal tissues. Cancer specimens from colorectal cancer patients were mechanically and enzymatically digested and separated into two fractions using a cell strainer: the organoid fraction (ORG), which was retained in a 40-?m strainer, and the flow-through fraction (FT), which passed through the strainer. The FT fraction mainly contained single cells and many of the cells with epithelial markers were dead. In contrast, the ORG contained irregular sheet- or tube-like structures, which were termed organoids. After overnight culture, the organoids became spherical and bright with a smooth surface and were termed cancer tissue–originated spheroids (CTOSs). During formation, the CTOSs were draped with cellular debris that easily detached with pipetting. The diameter of the CTOSs was ?40–500 ?m, depending on the cell strainer size. A CTOS with a diameter of 100 ?m consisted of ?100 cells. The authors were able to prepare CTOSs in most of the colorectal cancer specimens regardless of the disease stage and histology. CTOSs were highly viable with only a few dead cells on the outside. CTOSs could be cultured and expanded in vitro. Extracellular matrix was critical for favorable growth of CTOSs. Excellent proliferation was accomplished with the use of a 3D type I collagen-based culture system and serum-free medium designed for embryonic stem cells. The success rate for growth was 73.5%. CTOSs were composed of highly purified and viable cancer cells, that retained the characteristics of the parental tumors, grew in vitro and could be used for personalized diagnostic applications, including chemosensitivity assays.
This study demonstrated the usefulness of cancer tissue–originated spheroid (CTOS) method for the primary culture of lung cancer cells. Surgical specimens and pleural effusion samples from lung cancer patients were minced with a scalpel into approximately one square mm pieces, and washed with Hank’s balanced salt solution. Specimens were then digested in DMEM supplemented with Liberase DH, at 37°C for 1 to 2 hours with gentle stirring by a magnetic bar. Digested tissue suspensions were passed through 500-?m and 250-?m metal mesh filters to remove large masses of undigested fragments. Suspensions were further filtered through 100-?m and 40-?m cell strainers. Fragments on the cell strainer and cells in the flow-through fractions were collected separately, and were each washed with HBSS and cultured in StemPro hESC medium in a nontreated dish. Pleural effusions were transferred to 50-ml tubes and centrifuged at 200 g. Pellets were resuspended in HBSS, filtered through 40-?m cell strainers, and collected and cultured in the same manner as surgical specimens. CTOSs were embedded in Matrigel GFR and cultured in 100 ?l of basal medium containing growth factors (NRG1, Long-IGF1, bFGF , Activin A, or EGF). Basal medium consisted of DMEM/F12, 2% BSA fraction V, nonessential amino acids, penicillin, streptomycin, ascorbic acid, human transferrin, ?-mercaptoethanol, and trace elements. Lung carcinoma CTOSs grew in vitro and could be used for personalized chemosensitivity assays.
Chemical dissociation
Numerous types of cations (such as Ca2+and Mg2+) are used to maintain of cell surface integrity and the intracellular structural matrix [Li WC]. Chemical dissociation is a process where sequestration of these compounds from epithelial cells has been used to loosen intercellular bonds [Castell JV]. Sequestration is best achieved by exposure to EDTA or EGTA or complexes of tetraphenylboron plus potassium ions, which have been used to dissociate liver tissue, intestinal crypt cells and solid mammary tissue [ Harris CC]. Besides chelation, hypertonic solutions of disaccharides (sucrose, maltose, lactose, and cellobiose) have been reported to split gap junctions and zona occludentes, whose presence may be responsible for the clusters of cells that sometimes remain after enzymatic tissue digestion [Goodenough DA].
Mechanical dissociation
Mechanical dissociation of tissue involves repeated mincing with scissors or sharp blades, scraping the tissue surface, homogenization, filtration through a nylon or steel mesh (50 to 100 μm opening), vortexing, repeated aspiration through pipettes or sequentially smaller needles (e.g., 16-,20-, and 23-gage), application of abnormal osmolarity stress, or any combination of these techniques [Cunningham RE]. Usually, tumor specimens are first minced into small pieces (~1 mm) and then washed in tissue-specific medium to remove loosely bound cells or non-specific debris by gentle agitation. This process generates single cell suspension quickly with minimal number of steps. However, dissociation of tumor tissue using mechanical methods is not a suitable technique, for obtaining tumor cells for culture because the technique results in a high percentage of dead cells which secrete degrading enzymes
Accumulation of genetic aberrations in cancer cell lines occurs with increasing number of passages severely limiting their usefulness for personalized medicine Research reviewed here indicates that isolation and culturing of primary tumor cells in 3D is possible and it is likely that primary 3D tumor cultures will soon be routinely used for individualized solid tumor testing. Modern tissue repositories will need to be ready to support these activities as they expand from their traditional research role to active participation in personalized medicine.
Clearly Auctoresonline and particularly Psychology and Mental Health Care Journal is dedicated to improving health care services for individuals and populations. The editorial boards' ability to efficiently recognize and share the global importance of health literacy with a variety of stakeholders. Auctoresonline publishing platform can be used to facilitate of optimal client-based services and should be added to health care professionals' repertoire of evidence-based health care resources.
Journal of Clinical Cardiology and Cardiovascular Intervention The submission and review process was adequate. However I think that the publication total value should have been enlightened in early fases. Thank you for all.
Journal of Women Health Care and Issues By the present mail, I want to say thank to you and tour colleagues for facilitating my published article. Specially thank you for the peer review process, support from the editorial office. I appreciate positively the quality of your journal.
Journal of Clinical Research and Reports I would be very delighted to submit my testimonial regarding the reviewer board and the editorial office. The reviewer board were accurate and helpful regarding any modifications for my manuscript. And the editorial office were very helpful and supportive in contacting and monitoring with any update and offering help. It was my pleasure to contribute with your promising Journal and I am looking forward for more collaboration.
We would like to thank the Journal of Thoracic Disease and Cardiothoracic Surgery because of the services they provided us for our articles. The peer-review process was done in a very excellent time manner, and the opinions of the reviewers helped us to improve our manuscript further. The editorial office had an outstanding correspondence with us and guided us in many ways. During a hard time of the pandemic that is affecting every one of us tremendously, the editorial office helped us make everything easier for publishing scientific work. Hope for a more scientific relationship with your Journal.
The peer-review process which consisted high quality queries on the paper. I did answer six reviewers’ questions and comments before the paper was accepted. The support from the editorial office is excellent.
Journal of Neuroscience and Neurological Surgery. I had the experience of publishing a research article recently. The whole process was simple from submission to publication. The reviewers made specific and valuable recommendations and corrections that improved the quality of my publication. I strongly recommend this Journal.
Dr. Katarzyna Byczkowska My testimonial covering: "The peer review process is quick and effective. The support from the editorial office is very professional and friendly. Quality of the Clinical Cardiology and Cardiovascular Interventions is scientific and publishes ground-breaking research on cardiology that is useful for other professionals in the field.
Thank you most sincerely, with regard to the support you have given in relation to the reviewing process and the processing of my article entitled "Large Cell Neuroendocrine Carcinoma of The Prostate Gland: A Review and Update" for publication in your esteemed Journal, Journal of Cancer Research and Cellular Therapeutics". The editorial team has been very supportive.
Testimony of Journal of Clinical Otorhinolaryngology: work with your Reviews has been a educational and constructive experience. The editorial office were very helpful and supportive. It was a pleasure to contribute to your Journal.
Dr. Bernard Terkimbi Utoo, I am happy to publish my scientific work in Journal of Women Health Care and Issues (JWHCI). The manuscript submission was seamless and peer review process was top notch. I was amazed that 4 reviewers worked on the manuscript which made it a highly technical, standard and excellent quality paper. I appreciate the format and consideration for the APC as well as the speed of publication. It is my pleasure to continue with this scientific relationship with the esteem JWHCI.
This is an acknowledgment for peer reviewers, editorial board of Journal of Clinical Research and Reports. They show a lot of consideration for us as publishers for our research article “Evaluation of the different factors associated with side effects of COVID-19 vaccination on medical students, Mutah university, Al-Karak, Jordan”, in a very professional and easy way. This journal is one of outstanding medical journal.
Dear Hao Jiang, to Journal of Nutrition and Food Processing We greatly appreciate the efficient, professional and rapid processing of our paper by your team. If there is anything else we should do, please do not hesitate to let us know. On behalf of my co-authors, we would like to express our great appreciation to editor and reviewers.
As an author who has recently published in the journal "Brain and Neurological Disorders". I am delighted to provide a testimonial on the peer review process, editorial office support, and the overall quality of the journal. The peer review process at Brain and Neurological Disorders is rigorous and meticulous, ensuring that only high-quality, evidence-based research is published. The reviewers are experts in their fields, and their comments and suggestions were constructive and helped improve the quality of my manuscript. The review process was timely and efficient, with clear communication from the editorial office at each stage. The support from the editorial office was exceptional throughout the entire process. The editorial staff was responsive, professional, and always willing to help. They provided valuable guidance on formatting, structure, and ethical considerations, making the submission process seamless. Moreover, they kept me informed about the status of my manuscript and provided timely updates, which made the process less stressful. The journal Brain and Neurological Disorders is of the highest quality, with a strong focus on publishing cutting-edge research in the field of neurology. The articles published in this journal are well-researched, rigorously peer-reviewed, and written by experts in the field. The journal maintains high standards, ensuring that readers are provided with the most up-to-date and reliable information on brain and neurological disorders. In conclusion, I had a wonderful experience publishing in Brain and Neurological Disorders. The peer review process was thorough, the editorial office provided exceptional support, and the journal's quality is second to none. I would highly recommend this journal to any researcher working in the field of neurology and brain disorders.
Dear Agrippa Hilda, Journal of Neuroscience and Neurological Surgery, Editorial Coordinator, I trust this message finds you well. I want to extend my appreciation for considering my article for publication in your esteemed journal. I am pleased to provide a testimonial regarding the peer review process and the support received from your editorial office. The peer review process for my paper was carried out in a highly professional and thorough manner. The feedback and comments provided by the authors were constructive and very useful in improving the quality of the manuscript. This rigorous assessment process undoubtedly contributes to the high standards maintained by your journal.
International Journal of Clinical Case Reports and Reviews. I strongly recommend to consider submitting your work to this high-quality journal. The support and availability of the Editorial staff is outstanding and the review process was both efficient and rigorous.
Thank you very much for publishing my Research Article titled “Comparing Treatment Outcome Of Allergic Rhinitis Patients After Using Fluticasone Nasal Spray And Nasal Douching" in the Journal of Clinical Otorhinolaryngology. As Medical Professionals we are immensely benefited from study of various informative Articles and Papers published in this high quality Journal. I look forward to enriching my knowledge by regular study of the Journal and contribute my future work in the field of ENT through the Journal for use by the medical fraternity. The support from the Editorial office was excellent and very prompt. I also welcome the comments received from the readers of my Research Article.
Dear Erica Kelsey, Editorial Coordinator of Cancer Research and Cellular Therapeutics Our team is very satisfied with the processing of our paper by your journal. That was fast, efficient, rigorous, but without unnecessary complications. We appreciated the very short time between the submission of the paper and its publication on line on your site.
I am very glad to say that the peer review process is very successful and fast and support from the Editorial Office. Therefore, I would like to continue our scientific relationship for a long time. And I especially thank you for your kindly attention towards my article. Have a good day!
"We recently published an article entitled “Influence of beta-Cyclodextrins upon the Degradation of Carbofuran Derivatives under Alkaline Conditions" in the Journal of “Pesticides and Biofertilizers” to show that the cyclodextrins protect the carbamates increasing their half-life time in the presence of basic conditions This will be very helpful to understand carbofuran behaviour in the analytical, agro-environmental and food areas. We greatly appreciated the interaction with the editor and the editorial team; we were particularly well accompanied during the course of the revision process, since all various steps towards publication were short and without delay".
I would like to express my gratitude towards you process of article review and submission. I found this to be very fair and expedient. Your follow up has been excellent. I have many publications in national and international journal and your process has been one of the best so far. Keep up the great work.
We are grateful for this opportunity to provide a glowing recommendation to the Journal of Psychiatry and Psychotherapy. We found that the editorial team were very supportive, helpful, kept us abreast of timelines and over all very professional in nature. The peer review process was rigorous, efficient and constructive that really enhanced our article submission. The experience with this journal remains one of our best ever and we look forward to providing future submissions in the near future.
I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.
I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.
I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.
Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.
“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.
Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.
