Publications by authors named "Timothy J Hunt"

16 Publications

  • Page 1 of 1

Generation of three human induced pluripotent stem cell sublines (UCLAi004-A, UCLAi004-B, and UCLAi004-C) for reproductive science research.

Stem Cell Res 2021 Jul 24;54:102446. Epub 2021 Jun 24.

Department of Molecular, Cell and Developmental Biology, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA, USA; Molecular Biology Institute, University of California, Los Angeles, CA, USA. Electronic address:

Three induced pluripotent stem cell sublines (hiPSCs) were generated from human dermal human dermal fibroblasts (HDFs) derived from a human skin punch biopsy. The biopsy was donated from a woman with known infertility due to ovarian failure. The hiPSC sublines were created using Sendai virus vectors and were positive for markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA-4. Pluripotency was verified using PluriTest analysis and in vitro differentiation using Taqman Real-Time PCR assays for somatic lineage markers. This participant's monozygotic twin sister also donated a skin-punch biopsy, whose resulting hiPSC lines were published previously as a resource.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.scr.2021.102446DOI Listing
July 2021

Generation of three human induced pluripotent stem cell sublines (UCLAi005-A, UCLAi005-B and UCLAi005-C) for reproductive science research.

Stem Cell Res 2021 Jul 8;54:102409. Epub 2021 Jun 8.

Department of Molecular, Cell and Developmental Biology, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA USA; Molecular Biology Institute, University of California, Los Angeles, CA, USA. Electronic address:

We generated three human induced pluripotent stem cell (hiPSC) sublines from human dermal fibroblasts (HDF) (MZT05) generated from a skin biopsy donated from a previously fertile woman. The skin biopsy was broadly consented for generating hiPSC lines for biomedical research, including unique consent specifically for studying human fertility, infertility and germ cell differentiation. hiPSCs were reprogrammed using Sendai virus vectors and were subsequently positive for markers of self-renewal. Pluripotency was further verified using PluriTest analysis and in vitro differentiation was tested using Taqman Real-Time PCR assays. These sublines serve as controls for hiPSC research projects aimed at understanding the cell and molecular regulation of female fertility.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.scr.2021.102409DOI Listing
July 2021

Generation of six human induced pluripotent stem cell sublines (MZT01E, MZT01F, MZT01N and MZT02D, MZT02G and MZT02H) for reproductive science research.

Stem Cell Res 2021 03 27;51:102204. Epub 2021 Jan 27.

Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA, USA; Molecular Biology Institute, University of California, Los Angeles, CA, USA.

Six human induced pluripotent stem cell sublines (hiPSCs) were generated from human dermal fibroblasts (HDFs) derived from skin biopsies donated from monozygotic twin women wherein one woman had proven fertility and her sister was infertile due to ovarian failure. Three hiPSC sublines were created from each twin's HDFs. hiPSCs were reprogrammed using Sendai virus vectors and were subsequently positive for markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA-4. Pluripotency was further verified using PluriTest. We show here that the hiPSC lines created from the twins are equivalent in measures of pluripotency and self-renewal, despite their differential diagnosis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.scr.2021.102204DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8043044PMC
March 2021

Intrabone transplantation of CD34+ cells with optimized delivery does not enhance engraftment in a rhesus macaque model.

Blood Adv 2020 12;4(24):6148-6156

Laboratory of Transplantation Immunotherapy, Cellular and Molecular Therapeutics Branch, National Heart, Lung, and Blood Institute.

Intrabone (IB) injection of umbilical cord blood has been proposed as a potential mechanism to improve transplant engraftment and prevent graft failure. However, conventional IB techniques produce low retention of transplanted cells in the marrow. To overcome this barrier, we developed an optimized IB (OIB) injection method using low-volume, computer-controlled slow infusion that promotes cellular retention in the marrow. Here, we compare engraftment of CD34+ cells transplanted in a myeloablative rhesus macaque (RM) model using the OIB method compared with IV delivery. RM CD34+ cells obtained by apheresis were split equally for transduction with lentiviral vectors encoding either green fluorescent protein or yellow fluorescent protein reporters. Following conditioning, one marked autologous population of CD34+ cells was injected directly IB using the OIB method and the other was injected via slow IV push into the same animal (n = 3). Daily flow cytometry of blood quantified the proportion of engrafting cells deriving from each source. Marrow retention was examined using positron emission tomography/computed tomography imaging of 89Zirconium (89Zr)-oxine-labeled CD34+ cells. CD34+ cells injected via the OIB method were retained in the marrow and engrafted in all 3 animals. However, OIB-transplanted progenitor cells did not engraft any faster than those delivered IV and contributed significantly less to hematopoiesis than IV-delivered cells at all time points. Rigorous testing of our OIB delivery system in a competitive RM myeloablative transplant model showed no engraftment advantage over conventional IV infusion. Given the increased complexity and potential risks of IB vs IV approaches, our data do not support IB transplantation as a strategy to improve hematopoietic engraftment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1182/bloodadvances.2020003040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757001PMC
December 2020

Generation of three human induced pluripotent stem cell sublines (MZT04D, MZT04J, MZT04C) for reproductive science research.

Stem Cell Res 2019 10 16;40:101576. Epub 2019 Sep 16.

Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, CA, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, CA, USA; Molecular Biology Institute, University of California, Los Angeles, CA, USA. Electronic address:

We generated three human induced pluripotent stem cell (hiPSC) sublines from human dermal fibroblasts (HDFs) (MZT04) generated from a skin biopsy donated from a previously fertile woman. The skin biopsy was broadly consented for generating hiPSC lines for biomedical research, including unique consent specifically for studying human fertility, infertility and germ cells. hiPSCs were reprogrammed using Sendai virus vectors and were subsequently positive for markers of self-renewal including OCT4, NANOG, TRA-1-81 and SSEA-4. Pluripotency was further verified using teratomas and PluriTest. These sublines serve as controls for hiPSC research projects aimed at understanding the cell and molecular regulation of female fertility and infertility.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.scr.2019.101576DOI Listing
October 2019

Research Pearls: How Do We Establish the Level of Evidence?

Arthroscopy 2018 12;34(12):3271-3277

Heartland Orthopedic Associates, Alexandria, Minnesota, U.S.A.

Evidence-based medicine (EBM) guidelines were first introduced in 1986 and were defined as the conscientious, explicit, and judicious use of current best evidence in making decisions about the care of individual patients. The practice of EBM means integrating individual clinical expertise with the best available external clinical evidence from systematic research. Level of evidence (LOE) stratifies publications from Level I to Level V and provides the foundation for EBM. Three questions should be asked when an LOE is assigned to a scientific article: (1) What is the research question? (2) What is the study type? and (3) What is the hierarchy of evidence? In cases in which LOE is not appropriate or relevant (basic science and laboratory-based investigations), a clinical relevance statement should be used. Unfortunately, study quality is not assessed by the assigned hierarchy level. LOE and EBM have increased the number of investigations published with better levels of evidence. As authors, reviewers, editors, and publishers, we desire a system that is consistent, effective, and reliable. Fortunately, the system has proven to have all of those attributes with good interobserver and intra-observer values. The increase in investigations with higher LOEs allows for more frequent use of EBM.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2018.10.002DOI Listing
December 2018

TFAP2C regulates transcription in human naive pluripotency by opening enhancers.

Nat Cell Biol 2018 05 25;20(5):553-564. Epub 2018 Apr 25.

Department of Molecular, Cell and Developmental Biology, University of California Los Angeles, Los Angeles, CA, USA.

Naive and primed pluripotent human embryonic stem cells bear transcriptional similarity to pre- and post-implantation epiblast and thus constitute a developmental model for understanding the pluripotent stages in human embryo development. To identify new transcription factors that differentially regulate the unique pluripotent stages, we mapped open chromatin using ATAC-seq and found enrichment of the activator protein-2 (AP2) transcription factor binding motif at naive-specific open chromatin. We determined that the AP2 family member TFAP2C is upregulated during primed to naive reversion and becomes widespread at naive-specific enhancers. TFAP2C functions to maintain pluripotency and repress neuroectodermal differentiation during the transition from primed to naive by facilitating the opening of enhancers proximal to pluripotency factors. Additionally, we identify a previously undiscovered naive-specific POU5F1 (OCT4) enhancer enriched for TFAP2C binding. Taken together, TFAP2C establishes and maintains naive human pluripotency and regulates OCT4 expression by mechanisms that are distinct from mouse.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41556-018-0089-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5926822PMC
May 2018

My Kind of Town (Chicago Is): Content Collections Optimize Learning Related to the 2018 AANA Annual Meeting.

Arthroscopy 2018 04;34(4):1001-1004

The 2018 Arthroscopy Association of North America Annual Meeting represents an opportunity to deepen one's understanding of a wide variety of topics. Arthroscopy journal readers have diverse practices and interests, and the meeting is designed to accommodate individual needs. The constructivist learning theory provides that scholars learn in many different ways. Thus, to enrich your learning experience, selected recently published Arthroscopy articles are suggested to supplement material presented at the meeting. The articles are collated on our web site in Content Collections, to allow meeting participants to prepare and to allow those unable to attend to remain engaged. We offer suggestions and encourage readers to customize their own learning experience.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2018.02.018DOI Listing
April 2018

Editorial Commentary: Go Ahead and Repair That Shoulder Rotator Cuff Tear in Your Obese Patient: Just Be Prepared to Admit Them.

Authors:
Timothy J Hunt

Arthroscopy 2018 03;34(3):762-763

Arthroscopic rotator cuff repair in the obese patient offers functional outcomes and rates of complications comparable to those seen in nonobese patients. Future prospective studies with better methodology, as well as including larger numbers of severely obese patients with a body mass index of 40 or greater, will help to further elucidate if obesity truly affects outcomes in rotator cuff repair. In the meantime, be sure to consider admission of your obese rotator cuff repair patients.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2017.10.007DOI Listing
March 2018

PRDM14 is expressed in germ cell tumors with constitutive overexpression altering human germline differentiation and proliferation.

Stem Cell Res 2018 03 4;27:46-56. Epub 2018 Jan 4.

Department of Molecular, Cell and Developmental Biology, Los Angeles, CA 90095, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, Los Angeles, CA 90095, USA; University of California Los Angeles, Los Angeles, CA 90095, USA. Electronic address:

Germ cell tumors (GCTs) are a heterogeneous group of tumors occurring in gonadal and extragonadal locations. GCTs are hypothesized to arise from primordial germ cells (PGCs), which fail to differentiate. One recently identified susceptibility loci for human GCT is PR (PRDI-BF1 and RIZ) domain proteins 14 (PRDM14). PRDM14 is expressed in early primate PGCs and is repressed as PGCs differentiate. To examine PRDM14 in human GCTs we profiled human GCT cell lines and patient samples and discovered that PRDM14 is expressed in embryonal carcinoma cell lines, embryonal carcinomas, seminomas, intracranial germinomas and yolk sac tumors, but is not expressed in teratomas. To model constitutive overexpression in human PGCs, we generated PGC-like cells (PGCLCs) from human pluripotent stem cells (PSCs) and discovered that elevated expression of PRDM14 does not block early PGC formation. Instead, we show that elevated PRDM14 in PGCLCs causes proliferation and differentiation defects in the germline.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.scr.2017.12.016DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5858915PMC
March 2018

Editorial Commentary: Animal Trials Using Stem Cells to Enhance Anterior Cruciate Ligament Treatment Are Interesting. But, Why Not More About Us Humans?

Authors:
Timothy J Hunt

Arthroscopy 2018 01;34(1):343-344

Although hampered by heterogeneous studies with low methodologic quality and high risk for bias, the current literature demonstrates the potential for enhanced healing of anterior cruciate ligament injuries during the early phase using adult stem cells in animal trials. There are only 2 published controlled clinical trials on the subject matter, and they have small sample sizes, undefined cell numbers, and unstandardized selection criteria and surgical techniques. There is a clear need for studies with higher levels of evidence that would include long-term, larger animal studies ultimately leading to improved clinical trials.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2017.08.263DOI Listing
January 2018

A Chronic Cardiac Ischemia Model in Swine Using an Ameroid Constrictor.

J Vis Exp 2017 10 9(128). Epub 2017 Oct 9.

Animal Surgery and Resources Core, National Heart, Lung, and Blood Institute, National Institutes of Health;

Cardiovascular disease remains the number one cause of mortality in the United States. There are numerous approaches to treating these diseases, but regardless of the approach, an in vivo model is needed to test each treatment. The pig is one of the most used large animal models for cardiovascular disease. Its heart is very similar in anatomy and function to that of a human. The ameroid placement technique creates an ischemic area of the heart, which has many useful applications in studying myocardial infarction. This model has been used for surgical research, pharmaceutical studies, imaging techniques, and cell therapies. There are several ways of inducing an ischemic area in the heart. Each has its advantages and disadvantages, but the placement of an ameroid constrictor remains the most widely used technique. The main advantages to using the ameroid are its prevalence in existing research, its availability in various sizes to accommodate the anatomy and size of the vessel to be constricted, the surgery is a relatively simple procedure, and the post-operative monitoring is minimal, since there are no external devices to maintain. This paper provides a detailed overview of the proper technique for the placement of the ameroid constrictor.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3791/56190DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5752388PMC
October 2017

Sommelier Suggestions: The Arthroscopy Association of North America Annual Meeting Inspires a Content Collection Worth Tasting.

Arthroscopy 2017 May;33(5):885-887

The 2017 Arthroscopy Association of North America Annual Meeting Program inspires a Content Collection of Arthroscopy journal articles worthy of review. A foundation of a credible podium presentation is the published medical literature. Your Editors thus suggest recent publications that seem particularly relevant in the context of the 2017 annual meeting. Consider these articles as one would a suggestion for a good glass of wine to complement a delicious meal.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2017.03.003DOI Listing
May 2017

Editorial Commentary: The Time Has Come to Try Intra-articular Platelet-Rich Plasma Injections for Your Patients With Symptomatic Knee Osteoarthritis.

Authors:
Timothy J Hunt

Arthroscopy 2017 Mar;33(3):671-672

Platelet-rich plasma injections, in a systematic review and meta-analysis of 10 Level I randomized control trials, were found to provide more pain relief and better functional outcomes than hyaluronic acid in patients with knee osteoarthritis at 12 months after injection. The time has come for those of us who have not yet tried platelet-rich plasma injections in our patients with symptomatic knee osteoarthritis to do so.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2016.12.006DOI Listing
March 2017

Editorial Commentary: Posterior Cruciate Ligament Reconstruction--Do Not Abandon the C-Arm Quite Yet.

Authors:
Timothy J Hunt

Arthroscopy 2016 Mar;32(3):493-4

Accurate tibial tunnel placement using the arthroscopically-assisted anatomic fovea landmark technique in transtibial posterior cruciate ligament reconstruction is possible without the use of fluoroscopic imaging. However, until a prospective, randomized controlled trial comparing the C-arm and anatomic fovea landmark techniques is completed, abandonment of the C-arm in posterior cruciate ligament reconstruction cannot be recommended.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2015.12.024DOI Listing
March 2016

Editorial Commentary: Intra-articular Corticosteroid Injection at the Time of Knee Arthroscopy Is Not Recommended.

Authors:
Timothy J Hunt

Arthroscopy 2016 Jan;32(1):96

In a population of Medicare patients undergoing knee arthroscopy, a significant increase in the incidence of postoperative infection at 3 and 6 months was found in patients who received an intra-articular corticosteroid injection at the time of knee arthroscopy compared with a matched control group that did not receive an injection. Intra-articular corticosteroid injection at the time of knee arthroscopy is not recommended.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.arthro.2015.11.014DOI Listing
January 2016
-->