The question of this article is to determine if skin dermal papilla (DP) cells can differentiate into induced pluripotent stem cells (iPSCs) by using the transcription factor, OCT4, instead of using all four transcription factor, OCT4, Sox2, Klf4, and c-Myc, which are usually used to differentiate somatic cells into stem cells.
The scientists came up with the question because two out of the four transcription factors, Klf4 and c-Myc, are oncogenic gene, thereby it is best to replace these genes with other safer alternatives. Reprogramming cells into induced pluripotent stem cells impose risks because the process requires several transcription factors. To reduce the risks, programming iPSCs should require as little transcription factors
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Then for in vivo differentiation, two mice mated and all the blastocysts were collected. 1TF iPSCs injected into each blastocyst and the blastocysts were transplanted into the uterus of pseudopregnant female mice. 1TF chimeric mice were born and tested for gene expression. Then, 1TF mice mated to create F1 pup. These generations were used to test for indication of potential risks that occurred because of the differentiation.
In the results, the control group showed Oct4-GFP positive in five days while experimental group, 1TF iPSC, showed Oct4-GFP positive in 18 days. 1TF iPSC with only OCT4 as the transcription factor had reprogramming efficiency of 0.088%, which is higher than the control group that used all four transcription factors. 1TF iPS cell is morphology similar to ES cells 1TF iPSC colonies were stained positive for nanog, Oct4, Sox2, and SSEA-1. In addition, genes in expressed in the control group were all expressed in the experimental group. Like all pluripotent stem cells, 1TF iPSCs differentiated into the three germ layers, endoderm, mesoderm, and ectoderm. For in vivo, Oct4 were expressed in gonads of 1TF chimeric mice. 17 out of 43 mice grew into adult mice with the same agouti coat color as their parent. And in the third generation, the pup has no tumor formation, Oct4 -GFP was expressed as well as Lef1-RFP, which was the gene of the first generation Lef1-RFP/Oct4-GFP/Rosa26-lacZ transgenic
Pluripotent stem cells are the stem cells that can only differentiate into a limited range of differentiated cells. (2) They have the ability to give rise to all somatic cells from ectoderm, mesoderm and endoderm, as well as gametes. Naturally it can be found in embryos as Embryonic stem cells (ES cells). Induced pluripotent stem cells (iPS cells or iPSC) are the pluripotent stem cells that are generated directly from adult cells, first discovered by Shinya Yamanaka in 2006 by using a set of reprogramming factors (Oct4, Sox2, Klf4, and c-Myc or LIN28 and Nanog) (3) to reprogram mature cells back to a pluripotent state (4).
To give a short overview of the steps that will be taken to complete the study. Obtaining stem cells, whether adult, embryonic or induced, shall be done using healthy mouse models and after ethical approval has been gained. The process to derive them will be detailed below, however they are also purchasable commercially with the benefit of being well studied and accompanied by a detailed analysis of properties, however with a
The creation of induced pluripotent stem cells by direct reprogramming has allowed for the circumvention of using embryonic stem cells while still leaving the cells with the ability to maintain pluripotency. Instead of ES cells which were originally derived from the epiblast of mouse embryos, IPS cells were generated from mouse embryonic fibroblasts. This eliminated both any ethical concerns for whether those cells were a living being or not and the need to destroy embryos at the blastocyst stage. An advantage of IPS cells is that they are derived from human somatic cells which makes them easy to acquire due to the possibility of using skin or blood cells. They can also be grown and differentiated individually for each person that the sample of somatic cells is taken from which eliminates the possibility of having any immune reaction and rejection to the differentiated cells during transplantation. These characteristics of IPS cells are important because they are what enables us to safely and accurately transform these affected cells from patients cells into neurons and confidently study them.
Embryonic stem cells (hESC) are pluripotent. They are obtained from the inner mass of a 5-6 day old human blastocyst that consists of approximately 100 cells (Bongso & Lee, 2005, p. 3).
Human embryonic stem cells (hESCs) are pluripotent and are obtained from the inner mass of a 4-5 day old human blastocyst that consists of approximately 100 cells (“Stem cell research,” 2009).
Keywords: ethical, embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), disease, drug development, research
Those people also argue that society would begin to accept the concept or destroying one potential life in order to save another. The final source of embryonic stem cells is opposed only because people find creating an embryo only to destroy it later is morally wrong and inhumane. Those that are in favor or neutral to creating the embryos point out that it is acceptable for an embryo that is created accidentally to be destroyed as a consequence of pregnancy, but to create an embryo with the intent to use the stem cells it is composed of to save a life is wrong (Hug). The second type of stem cell are pluripotent stem cells. Pluripotent stem cells come from fetal tissues instead of the embryo itself. Fetal tissues include the umbilical cord and the blood inside it, and the placenta. There is also a second kind of pluripotent stem cell. These stem cells are called induced pluripotent stem cells, or iPSCs. iPSCs are stem cells were adult stem cells, but have been reprogrammed into their embryonic stages. Though iPSCs resemble human embryonic stem cells, they are not usually accepted into the body, and so they are introduced to other adult stem cells with a virus that has caused some cancers to develop (National Institutes of Health).
The ability to manipulate the stem cell corresponding to a specific organ/tissue remains important. A type of stem cell that can be manipulated is the embryonic stem cell. These stem cells descend from embryos aging from three to five days (Watt) (Driskell). During earlier stages, scientists describe embryonic stem cells as “blastocysts” which contain over one-hundred and fifty cells (Watt) (Driskell). They duplicate into more cells or transform to any cell located in the body (Watt) (Driskell). This “duplication” allows embryonic stem cells to regenerate and repair diseased tissues. Embryonic stem cells gain importance in cancer treatments—if doctors diagnose patients with leukemia, then during chemotherapy, the doctor can infuse embryonic stem cells into the body. Since the cells are young, they can repair the targeted cell, aiding cancer treatments and the patient. In addition, this technique is used with another type of embryonic stem cell called “pluripotent stem cells”. Pluripotent stem cells originate as inner mast cells (cells
The human embryo for embryonic stem cell research requires the ova from a woman to make this possible. This requires many risks to the woman giving the egg. “Embryonic stem cells are pluripotent cells positioned in the early embryo” (Miller Ph.D., Levine Ph.D.). Pluripotent means that the cells are capable of developing into most of the body’s cell types and have the ability to aid and cure diseases. (Miller Ph.D., Levine Ph.D.). This pluripotency is what distinguishes between embryonic and adult stem cells. The embryonic stem cells can be generated in every cell type in the body and can indefinitely create themselves making it possible for tissue replacement in addition to finding cures for diseases. “Embryonic stem cells are human embryos that develop after fertilization into a blastocyst” (Miller Ph.D., Levine Ph.D.). Hundreds of immune system diseases and rare genetic disorders are believed to be among the possible to be aided or cured using embryonic stem cells. Embryonic stem cells
This paper aims to identify key developments in the evolution of indued pluripotent stem cells and how these developments will impact the medical field. Beginning with a comprehensive exploration of the history and discovery of stem cells, it will highlight the challenges historically faced by researchers and medical professionals prior to the discovery of defined factors in adult cells. Using published research for reference, it will describe the process of discovery and modern application of induced pluripotent stem cells, leaning heavily on the original work of Shinya Yamanaka of Kyoto University, who was awarded a Nobel Prize for her role in discovering adult pluripotency. Finally, it will take a forward perspective to predict how this technology may be used in the future of medicine and discuss some of the most controversial ethical questions regarding these potential uses.
iPSCs are adult stem cells that have been genetically reprogrammed to behave like the pluripotent stem cells found in embryos, i.e. can differentiate into any cell type in the human body. This was first completed successfully in mice in 2006 by Shinya Yamanaka and his team (Takahashi et al., 2006), then in humans in 2007 both by Yamanaka (Takahashi et al., 2007), and by James Thomson and his team in America independently (Yu, et al., 2007). Yamanaka and Thomson’s methods were similar. In the report by Yu et
In the research that will be conducted at Harvard, if permission is granted, the growth of the blastocyst would be stopped and it would not be implanted into a woman’s womb. Stem cells would be extracted for study, destroying the embryo.
I have worked at the broadband industry’s leader, Incognito Software, for over 10 years. Throughout this time I’ve created several innovations. Some standouts would be the company’s online customer knowledgebase, a new-hire onboarding program, and a quarterly technical newsletter. I am quite proud of one innovation: The Boot Camp at the annual Incognito User Conference.
To achieve these objectives, we will use three different mouse embryonic stem (ES) cell lines (R1, D3, E14), already present in the lab. In ongoing studies, it was noted that these cells presented all five of the ING genes with altered expression when they were induced to differentiate. We will first study if altering ING5 and ING4 expression using siING5 and siING4 to decrease it and pCI-ING5, pCI-ING4 to increase levels, will affect differentiation and the self-renewal rate of these lines. We will use sphere formation assays to assess self-renewal. We will also induce differentiation with these different expression constructs, thus measuring the influence of these proteins in the process. To determine the degree of differentiation with varying expression of the ING genes, we will use flow cytometry to follow changes in stem cells markers, such as OCT4, OLIG2 and Nestin16.
During the teenage years is when the human brain goes through the most drastic changes, both at the cellular level and at the emotional level. Teenage brains go through the most emotional distress because their frontal cortex is not fully developed. According to dr. Charles Nelson who was interviewed for the film; Inside The Teenage Brain and said, “...and because the child - the 13 or 14 or 15-year-old - still has an immature frontal cortex, they often do not make the most responsible, reasoned decisions.” This is one plausible explanation to most teenage attitude. Dr. Nelson also referred to mood swings, “But we think the ultimate responsibility for regulating these mood changes resides in the frontal cortex, and that's what's overseeing