Mouse Embryonic Fibroblasts (MEFs) are a type of fibroblast prepared from mouse embryo. MEFs show a spindle shape when cultured in vitro , a typical feature of fibroblasts. The MEF is a limited cell line. After several transmissions, MEFs will senesce and finally die off. Nevertheless, researchers can use several strategies, like virus infection or repeated transmission to immortalize MEF cells, which can let MEFs grown indefinitely in spite of some changes in characters. [1] [2]
MEFs are widely used in life science researches, especially in stem cell biology.
To prepare MEFs, pregnant female mice are needed. After killing the female mouse, the researcher should incise its stomach and then detach the embryo from the placenta in a Biosafety Cabinet. Then the liver and head should be taken out. Finally digest the remains by enzymes to obtain single isolated cells and culture the cells in a tissue culture dishes. MEF cells can be cultured in vitro in DMEM medium with 10% FBS. To transmit MEFs, researches should use trypsin to digest the cells (making them detach from the surface) and transmit 1/5 cells digested into a new dish. [3] [1]
In 1962, George Todaro and Howard Green, two researchers in New York University, immortalized MEFs by repeated transmission. These cells developed into the commonly used cell line NIH 3T3. [4]
MEFs treated by mitomycin or gamma rays (such treatment makes MEF stop mitosis) are widely used as feeder in embryonic stem cell culture because they can mimic the microenvironment in embryo. [5] In 2006, Shinya Yamanaka reprogrammed MEFs into iPSCs by introducing 4 factors, which is remarkable in the development of stem cell biology. [6]
In multicellular organisms, stem cells are undifferentiated or partially differentiated cells that can change into various types of cells and proliferate indefinitely to produce more of the same stem cell. They are the earliest type of cell in a cell lineage. They are found in both embryonic and adult organisms, but they have slightly different properties in each. They are usually distinguished from progenitor cells, which cannot divide indefinitely, and precursor or blast cells, which are usually committed to differentiating into one cell type.
A fibroblast is a type of biological cell typically with a spindle shape that synthesizes the extracellular matrix and collagen, produces the structural framework (stroma) for animal tissues, and plays a critical role in wound healing. Fibroblasts are the most common cells of connective tissue in animals.
Embryonic stem cells (ESCs) are pluripotent stem cells derived from the inner cell mass of a blastocyst, an early-stage pre-implantation embryo. Human embryos reach the blastocyst stage 4–5 days post fertilization, at which time they consist of 50–150 cells. Isolating the inner cell mass (embryoblast) using immunosurgery results in destruction of the blastocyst, a process which raises ethical issues, including whether or not embryos at the pre-implantation stage have the same moral considerations as embryos in the post-implantation stage of development.
Embryoid bodies (EBs) are three-dimensional aggregates formed by pluripotent stem cells. These include embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC)
Oct-4, also known as POU5F1, is a protein that in humans is encoded by the POU5F1 gene. Oct-4 is a homeodomain transcription factor of the POU family. It is critically involved in the self-renewal of undifferentiated embryonic stem cells. As such, it is frequently used as a marker for undifferentiated cells. Oct-4 expression must be closely regulated; too much or too little will cause differentiation of the cells.
Sir Martin John EvansFLSW is an English biologist who, with Matthew Kaufman, was the first to culture mice embryonic stem cells and cultivate them in a laboratory in 1981. He is also known, along with Mario Capecchi and Oliver Smithies, for his work in the development of the knockout mouse and the related technology of gene targeting, a method of using embryonic stem cells to create specific gene modifications in mice. In 2007, the three shared the Nobel Prize in Physiology or Medicine in recognition of their discovery and contribution to the efforts to develop new treatments for illnesses in humans.
A stem cell line is a group of stem cells that is cultured in vitro and can be propagated indefinitely. Stem cell lines are derived from either animal or human tissues and come from one of three sources: embryonic stem cells, adult stem cells, or induced pluripotent stem cells. They are commonly used in research and regenerative medicine.
The stem cell controversy concerns the ethics of research involving the development and use of human embryos. Most commonly, this controversy focuses on embryonic stem cells. Not all stem cell research involves human embryos. For example, adult stem cells, amniotic stem cells, and induced pluripotent stem cells do not involve creating, using, or destroying human embryos, and thus are minimally, if at all, controversial. Many less controversial sources of acquiring stem cells include using cells from the umbilical cord, breast milk, and bone marrow, which are not pluripotent.
SOX genes encode a family of transcription factors that bind to the minor groove in DNA, and belong to a super-family of genes characterized by a homologous sequence called the HMG-box. This HMG box is a DNA binding domain that is highly conserved throughout eukaryotic species. Homologues have been identified in insects, nematodes, amphibians, reptiles, birds and a range of mammals. However, HMG boxes can be very diverse in nature, with only a few amino acids being conserved between species.
3T3 cells are several cell lines of mouse embryonic fibroblasts. The original 3T3 cell line was established in 1962 by two scientists then at the Department of Pathology in the New York University School of Medicine, George Todaro and Howard Green. Todaro and Green originally obtained their 3T3 cells from Swiss albino mouse embryo tissue. Later, as a principal investigator position at the National Cancer Institute in Bethesda, Maryland, Todaro repeated the isolation procedure from the NIH Swiss mouse embryo with his students and established NIH-3T3 cell line.
The inner cell mass (ICM) or embryoblast is a structure in the early development of an embryo. It is the mass of cells inside the blastocyst that will eventually give rise to the definitive structures of the fetus. The inner cell mass forms in the earliest stages of embryonic development, before implantation into the endometrium of the uterus. The ICM is entirely surrounded by the single layer of trophoblast cells of the trophectoderm.
Induced pluripotent stem cells are a type of pluripotent stem cell that can be generated directly from a somatic cell. The iPSC technology was pioneered by Shinya Yamanaka and Kazutoshi Takahashi in Kyoto, Japan, who together showed in 2006 that the introduction of four specific genes, collectively known as Yamanaka factors, encoding transcription factors could convert somatic cells into pluripotent stem cells. Shinya Yamanaka was awarded the 2012 Nobel Prize along with Sir John Gurdon "for the discovery that mature cells can be reprogrammed to become pluripotent."
Shinya Yamanaka is a Japanese stem cell researcher and a Nobel Prize laureate. He is a professor and the director emeritus of Center for iPS Cell Research and Application, Kyoto University; as a senior investigator at the UCSF-affiliated Gladstone Institutes in San Francisco, California; and as a professor of anatomy at University of California, San Francisco (UCSF). Yamanaka is also a past president of the International Society for Stem Cell Research (ISSCR).
3T3-L1 is a sub clonal cell line derived from the original 3T3 Swiss albino cell line of 1962. The 3T3 original cell line was isolated from a mouse embryo and propagated for this specific line of 3T3 cells is used to study adipose tissue-related diseases and dysfunctions. The 3T3-L1 Swiss subclone line has been widely utilized, since its development, due to its affinity for lipid droplet deposition in vitro. 3T3-L1 cells have a fibroblast-like morphology, but, under appropriate conditions, the cells differentiate into an adipocyte-like phenotype, providing an exemplar model for white adipocytes. 3T3-L1 cells can be utilized to study a number of cellular and molecular mechanisms related to insulin-resistance, obesity, and diabetes in vitro. Aside from its usages, this cell line is widely developed and can be purchased for continuous propagation for numerous research studies. 3T3-L1 cells of the adipocyte morphology increase the synthesis and accumulation of triglycerides and acquire the signet ring appearance of adipose cells. These cells are also sensitive to lipogenic and lipolytic hormones, as well as drugs, including epinephrine, isoproterenol, and insulin.
Embryomics is the identification, characterization and study of the diverse cell types which arise during embryogenesis, especially as this relates to the location and developmental history of cells in the embryo. Cell type may be determined according to several criteria: location in the developing embryo, gene expression as indicated by protein and nucleic acid markers and surface antigens, and also position on the embryogenic tree.
Cell potency is a cell's ability to differentiate into other cell types. The more cell types a cell can differentiate into, the greater its potency. Potency is also described as the gene activation potential within a cell, which like a continuum, begins with totipotency to designate a cell with the most differentiation potential, pluripotency, multipotency, oligopotency, and finally unipotency.
The STAT3-Ser/Hes3 signaling axis is a specific type of intracellular signaling pathway that regulates several fundamental properties of cells.
Directed differentiation is a bioengineering methodology at the interface of stem cell biology, developmental biology and tissue engineering. It is essentially harnessing the potential of stem cells by constraining their differentiation in vitro toward a specific cell type or tissue of interest. Stem cells are by definition pluripotent, able to differentiate into several cell types such as neurons, cardiomyocytes, hepatocytes, etc. Efficient directed differentiation requires a detailed understanding of the lineage and cell fate decision, often provided by developmental biology.
OP9 cells are a cell line derived from mouse bone marrow stromal cells (mesenchyme). These cells are now characterized as stem cells. When co-cultured with embryonic stem cells (ESC), OP9 cells can induce ESC to differentiate into blood cells by serving as a feeder layer. They have the potential to be used in cell therapy, regenerative medicine and as immunomodulators.
ATPase Domain 3B (ATAD3B) is a protein that in humans is encoded by the ATAD3B gene. ATAD3 is part of the AAA protein family. The function of ATAD3B is not yet well understood by the scientific community. In humans the gene is located at 1p36.33.
{{cite journal}}
: CS1 maint: numeric names: authors list (link)