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Stem cell technology

Stem cell technology

Technology used for isolating, generating, growing and controlling the differentiation of stem cells into specialized cells and the application of stem cell technology for medical and non-medical purposes such as producing meat through cellular agriculture.


Stem cells are self-renewing cells that have the potential to become multiple different cell types in the body. Embryonic stem cells are isolated from developing embryos. Adult stem cells are isolated from blood or tissues. Induced pluripotent stem (iPS) cells, also abbreviated iPSCs, are a type of stem cell derived from adult cells that are reprogrammed in the lab.

Stem cell culture

Stem cell culture shares many of the same protocols and equipment as standard mammalian cell culture but there are special conditions needed to maintain them in an undifferentiated state such as supplying certain growth factors and growing cells in colonies on a feeder cell layer. While mouse-derived feeder cells are often used in research, human feeder cells (Xeno-free) or feeder-free culture systems are needed for clinical use to avoid patient exposure to animal pathogens. Stem cells are also used to generate cultured meat, a product that also aims to be grown feeder-free and serum-free to avoid relying on animal use. The Cultured Meat Foundation is aiming to create a bank of induceded pluripotent cells (iPS cells) from cow, pig, chicken, tuna, salmon and lobster that are grown serum free and feeder free.

Controlling stem cell differentiation.

In development stem cells receive cues from their environment causing them to up regulate and down regulate genes which control stemness and cell differentiation into specialized cell types. In culture, stem cell scientists and engineers seek to program cells in a predictable way by supplying growth factors, physical cues or modifying the expression of genes that control stemness or key cell type specific genes.

Tracking stem cells and their differentiation in culture
Tracking stem cells in living tissues
  • Direct Labeling. A labeling agent is introduced into the cells, which is incorporated or attached to the cells prior to transplantation. Radionuclides are used for direct labeling but allow only short-term monitoring because radiodecay and diffusion of signal though cell division and dispersion. Other direct labelling materials include nanoparticles or quantum dots. Depending on the radiotracer, imaging can be performed using single-photon emission computed tomography (SPECT) or positron emission tomography (PET). Magnetic resonance imaging (MRI) can track cells labeled with superparamagnetic iron oxide particles (SPIO).
  • Indirect Labeling. Several reporter gene strategies have been used in pre-clinical stem cell transplant studies to study the long-term fate of transplanted cells. A reporter gene may be introduced into cells prior to transplantation that expresses fluorescent proteins such as green fluorescent protein or bioluminescence. The herpes simplex virus type 1 thymidine kinase (HSV1-tk) system phosphorylates an exogenously administered substrate. PET imaging can be used to detect a dopamine 2-like receptor reporter gene which codes for a cell membrane protein that binds an exogenously administered probe. Another reporter gene system uses a thyroid transmembrane protein, sodium-iodide symporter (NIS) which transports iodine into cells in exchange for sodium, which can be imaged with PET or SPECT with radioactive iodine or Technetium Tc-99m pertechnetate. MRI detected reporter genes are generally iron homeostasis proteins, reporter enzymes and reporter genes which generate chemical exchange saturation transfer. Reporter gene systems are not used in humans due to the potential for adverse effects of integration of foreign DNA into human cells. Safe methods for long-term monitoring of transplanted cells in human patients are needed.
  • Single-cell RNA sequencing
Stem cell isolation
  • Pre-plating makes use of the phenomenon that stem cells tend to adhere to culture plates and dishes more than the rest of the cells in a population. Pre-plating has been shown to enrich for mesenchymal stem cells (MSCs) from bone marrow or human adipose-derived stem cells from lipoaspirate from liposuction procedure.
  • Density gradient centrifugation. Separation of cells by density gradient requires knowledge of the density of the target cell type. A density gradient is established in a test tube and centrifuged cells accumulate at a position where density of cells matches the density of medium. Ficoll-paque, Percoll and RosetteSep are used as media for stem cell separation by density gradient.
Generating induced pluripotent stem (iPS) cells

Induced Pluripotent Stem Cells, abbreviated as iPS cells or iPSCs, are generated in the laboratory by treating adult differentiated cells from sources such as skin, with factors that reprogram the cells. The pluripotent cells can then be treated with factors that signal the cells to differentiate into another cell type.

Cell delivery
Stem cell standards and oversight
Stem cell characterization
Stem cell applications in medicine
Tissue Engineering

Tissue engineering is the differentiation of cells into specific cell types using cues like growth factors, physical scaffolds and other 3D cell culture approaches to produce functional replacement tissue or organs. Synthetic biology approaches in tissue engineering are being used for programming cells to self assemble into tissues.

Cell therapy

Hematopoietic stem cell transplants are routinely used to treat patients with cancers and other blood and immune system disorders. Hematopoietic cells can come from bone marrow, blood or umbilical cords. Many cell therapies use MSCs (mesenchymal stromal cells/mesenchymal stem cells) or PBSC (Peripheral blood stem cell) or other stem cells which may be effective due to their ability to differentiate into various tissues or due to secretion of paracrine factors such as growth factors, anti-inflammatory or pro-angiogenic factors. Cells may also be used as carriers of therapeutic agents.

Immunotherapy with engineered cells

Cell therapies being used or in development for treating cancer or autoimmune diseases use T cells and regulatory T cells (Tregs) respectively engineered with chimeric antigen receptors (CARs). CAR-T and TCR-T are engineered T cells and CAR-Treg and TCR-Treg therapies are engineered Tregs. FDA-approved CAR-T products are generated from harvesting autologous T cells from patients, followed by gene editing and infusion back into the patients. Allogeneic or universal cell therapy products, also called off-the-shelf cell therapy products would allow a broader implementation of these cell therapies. One method being researched is to derive natural killer (NK) cells from iPSCs and engineer iPSC-derived NK cells to target and kill cancer cell similarly to CAR-T cells.

Platelets carrying immunotherapy agents such as antibodies against immune checkpoint protein PD-1 have been tethered to hematopoietic stem cells to bring this immunotherapy into the bone marrow where it is needed to treat acute myeloid leukemia (AML). The technology has been developed in the lab of Zhen Gu, Founder of Zencapsule and Professor of bioengineering at UCLA Samueli School of Engineering.

Companies developing cell therapies

Amyotrophic lateral sclerosis (ALS)


  • RHEACELL manufactures GMP-compliant limbal stem cells from cornea tissue that are ABCB5+, a marker associated with clinical success in transplantation of these cells for treating blindness

Bone and cartilage repair and wound care

Bone marrow transplants


  • ZenCapsule, focused on developing delivery strategies for cancer immunotherapy
  • Ziopharm Oncology

Crohn's disease

  • Alofisel, MSC product


Graft verus host disease (GvHD)

Kidney disease

Neurodegenerative disease

Cell therapy companies that target multiple diseases

Cells engineered to carry biotherapeutics
Gene therapy

While one approach to gene therapy is to deliver genes or gene editing components in vivo, the other approach is to transduce the therapeutic gene or gene editing components into patient-derived cells ex vivo. Then the patient’s gene edited or genetically engineered cells are transplanted back into the patient. Hematopoetic stem cells and iPS cells are often used for this approach.

Disease modeling, drug development and drug testing

Stem cells are used to grow organoids and organ-on-a-chip devices for disease modeling, drug development, drug testing and personized medicine.

Extracellular vesicles derived from stem cells

The secretome is the set of molecules such as proteins, nucleic acids, lipids and extracellular vesicles secreted to the extracellular space. Microvesicles and exosomes are two classes of extracellular vesicles. Exosomes are smaller and are released by endosome fusion with the plasma membrane and microvesicles are shed from the plasma membrane. Exosomes were once thought to be a method for cells to discard unwanted proteins but are now known to have a role in intercellular communication with both neighboring and distant cells in the body.

Extracellular vesicles can be isolated from cell culture media and from body fluids. Since the regenerative effects of MSCs are partly mediated by secreted exosomes which carry proteins, ncRNA, RNA and lipids, MSC exosomes are being explored as a cell-free therapy in regenerative medicine.

Conditioned medium or cell culture medium previously used to grow MSCs and other stem cells could be used to extract therapeutic components. Conditioned medium derived from MSCs has been shown to be as effective as MSCs in many animal models for human disease.

Stem cell therapies aimed at treating injured myocardium in patients that had experienced heart failure were intended to differentiate into cardiomyocytes, engraft and integrate with the host tissue. The stem cells rarely differentiated into heart muscle and integrated into the host tissue, cardiac function was restored, due secretion of factors from the exosomes of the transplanted induce pluripotent cell (iPSC) derived cardiomyocytes.

Aegle Therapeutics - isolates extracellular vesicles from bone marrow derived MSCs to treat dermatological conditions

Exopharm - developing therapies based on exosomes from platelets and stem cells

Regeneus - clinical stage regenerative medicine company using MSCs and their secretions to treat knee osteoarthritis

Cell manufacturing for research and cell therapy

In contrast to drugs, which are chemically defined products that can be replicated, cell therapy and tissue engineering products are defined by their process. In order to standardize cell products the process needs to be well defined. Production may be scaled up so that the cell product is manufactured in one or two facilities and then shipped to locations where they are needed. Alternatively, production may be scaled out to multiple facilities such as dedicated hospital clean rooms, where it is produced in smaller batches on site.

Extracellular vesicle manufacturing
  • Stem Cell Exomere Program, a partnership between RoosterBio and Exopharm to develop and implement a standardized, scalable, commercially-viable biomanufacturing process and a cGMP-compliant Exomere product (exosomes) for regenerative medicine applications.
  • Exopharm
  • VivaZome Therapeutics
  • Lonza
Companies providing supplies for stem cell culture
Non-profit organizations


Further Resources


Essentials of Stem Cell Biology | ScienceDirect

Robert Lanza and Anthony Atala



Handbook of Stem Cells | ScienceDirect

Robert Lanza and Anthony Atala



Stem Cells

Mary Clarke, Jonathan Frampton



Stem Cells - 3rd Edition

Christine Mummery


January 20, 2021


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