Single-cell isolation allows genomic and transcriptomic analysis of one individual cell. It is also required to build a monoclonal cell line from rare cell isolation, that could be for example CRISPR-CAS9 gene edited cells. There are 3 popular methods: serial dilution, micro-manipulation and flow cytometry. None of these are easy or simple, and they often require expertise and experience. Fortunately, a new solution for everybody has come! It’s designed to isolate a single cell in a few seconds, and it’s (quite appropriately) called the Smart Aliquotor. [Read more…]
Tips for primary cell culture
Functionality and viability of primary cells can be impaired by incorrect thawing procedures, storage or culture conditions. It’s generally admitted that applying the same protocols as for cell lines leads will lead to bad cell quality. Well, based on our experience with primary cells, here are a few tips you can follow to ensure you get the best performance.
Simple and effective CRISPR CAS9 gene editing for primary cells
Vector-free CRISPR-CAS9 gene editing to accelerate therapeutic applications
A few years ago, Ayal Hendel et al (doi:10.1038/nbt.3290) published results revealing that chemical alterations to sgRNA enhance gene editing in primary cells. To demonstrate this, Matthew H Porteus’s team chose the targeted genes CCR5, HBB and IL2RG respectively involved in anti-HIV clinical trials, cell anaemia and thalassemia, and severe combined immunodeficiency. More recently, the same team tested several modified CAS9 mRNA. You can find the practical results on this poster introduced at the ASGC. [Read more…]
3 new types of Gastrointestinal (GI) Epithelial primary cells
Gastrointestinal (GI) primary cells represent a powerful approach for the in vitro study of the physiopathology of this unique tissue. Researchers have now identify intimate interactions between GI cells and its microbia with neurological disorders pointing out the need to access highly characterized sources of GI primary cells to design reliable and more physiologically relevant in vitro cellular models.
[Read more…]
Human Islets for Research – Which factors affect their quality?
The islets of Langerhans are the regions of the pancreas that contain its endocrine (i.e., hormone-producing) cells. Discovered in 1869 by German pathological anatomist Paul Langerhans, the islets of Langerhans constitute approximately 1% to 2% of the mass of the pancreas. There are about one million islets distributed throughout the pancreas of a healthy adult human. Each is separated from the surrounding pancreatic tissue by a thin fibrous connective tissue capsule. The islets of Langerhans contain beta cells, which secrete insulin, and play a significant role in diabetes.
Islets are widely used for transplantation to restore beta cell function from diabetes, offering an alternative to a complete pancreas transplantation or an artificial pancreas. Because the beta cells in the islets of Langerhans are selectively destroyed by an autoimmune process in type 1 diabetes, islet transplantation is a means of restoring physiological beta cell function in patients with type 1 diabetes.
Human Islets for Research (HIR)® are primary human islets processed from organ donor pancreases that have been approved for research but not for clinical transplantation of either the pancreas or the isolated islets. HIR® are obtained in a proprietary process of pancreas digestion and islet purification that results in uniformly high quality HIR® for delivery to diabetes investigators. Quality Control (QC) testing is routinely performed prior to release to assure uniform quality and function of these islets available for research. [Read more…]
“Extraordinary” CAR T Cells eliminate cancer after stem cell transplant
Donor-derived lymphocytes attack a patient’s cancer — but may attack the patient as well. Donor chimeric antigen receptor (CAR) T cells, on the other hand, brought remissions without this troubling complication.
CAR T cells continue to make waves: At the latest annual meeting of the American Association for the Advancement of Science, it was announced that CAR T cells, in which T cells from a patient are genetically reprogrammed to target cancer cells, removed all traces of cancer in the bone marrow of 27 out of 29 acute lymphoblastic leukemia patients. Nineteen of 30 individuals with non-Hodgkin lymphoma also responded in the form of partial or complete responses. The cumulative successes found in CAR T cell studies have labeled them “extraordinary.”[1] [Read more…]
Mimic human pulmonary function in vitro
If you’re studying human pulmonary function and pathophysiology, you need access to validated, highly characterised, human airway cellular models. A large collection of these cells is available for applications in asthma, inhalation toxicology and pulmonary inflammatory response. Let’s take a look at a selection of different pulmonary cell types I would advise, to boost your in vitro research in this area.
Cryopreserved Leukopaks: a solution for Cell Therapy Logistics
Human cells are critical raw materials for research and manufacturing of cell therapy products. However, access to freshly procured cells can be limited, creating a crucial need for a suitable alternative to fresh cells that are viable and functional — especially when transporting materials globally.
At tebu-bio, we chose HemaCare to investigate the viability and functionality of lymphocytes, both fresh and cryopreserved, from leukopaks (leukapheresis collections) procured within their FDA registered cGMP donor collection facility. [Read more…]
New in vitro cellular model for human Lipodystrophy
Lipodystrophies are disorders characterized by complete or selective loss of adipose tissue from various regions of the body. They might lead to severe metabolic disorders. The development of reliable cellular experimental models mimicking such diseases in vitro is extremely challenging (1). One of the main hurdle in the design of such in vitro cellular models is the access to reliable sources of well-qualified primary cells and the identification of optimal cell culture conditions.
Enriched Tregs for a multitude of research applications
When it comes to the body’s natural defenses, is it possible to have “too much of a good thing”? Absolutely. To spare the host, the immune system needs to distinguish it from the real enemy –infectious agents. Autoimmune disorders occur when this recognition frays and the body’s own cells and tissues are damaged.
That’s where regulatory T cells, or Tregs, come in. [Read more…]