![Optimized Induction of Human Hemogenesis in Adult Fibroblasts by Defined Factors and Hematopoietic Co-culture için kapak resmi Optimized Induction of Human Hemogenesis in Adult Fibroblasts by Defined Factors and Hematopoietic Co-culture için kapak resmi](/client/assets/d79c3e4af2b6d196/ctx/images/no_image.png)
Optimized Induction of Human Hemogenesis in Adult Fibroblasts by Defined Factors and Hematopoietic Co-culture
Başlık:
Optimized Induction of Human Hemogenesis in Adult Fibroblasts by Defined Factors and Hematopoietic Co-culture
Yazar:
Daniel, Michael Guillermo, author.
ISBN:
9780438030398
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (152 pages)
Genel Not:
Source: Dissertation Abstracts International, Volume: 79-10(E), Section: B.
Advisors: Kateri A. Moore Committee members: Nicole Dubois; Saghi Ghaffari; Eirini Papapetrou; Shahin Rafii; Michael Rendl.
Özet:
The inability to culture hematopoietic stem cells (HSCs) in vitro remains a significant problem in studying HSC biology. This dilemma hampers study of various hematologic pathologies, as well as disease modeling and drug testing efforts to understand and eventually treat these disorders. Recent work focuses on reprogramming pluripotent stem cells or somatic cells to generate HSCs, with most success found in those that attempt to follow known details of developmental hematopoiesis. Most studies, however, fall short of generating bona fide HSCs due to an incomplete understanding of hematopoiesis and the microenvironmental signals required for HSC induction and maintenance. Overexpression of the transcription factors (TFs) Gata2, Gfi1-b, and cFos (GGF) in mouse fibroblasts induces a transition through endothelial intermediates that gives rise to clonogenic CD45+ hematopoietic cells upon prolonged culture. Similar studies show that the same factors will reprogram human fibroblasts to hemogenic cells but their yield and functional potential requires improvement. We identified GFI1 as a TF that significantly improves the yield of functional HSC-like cells when in concert with GGF (herein named 3GF for GATA2, GFI1, GFI1B, and FOS) that possess a cell surface phenotype highly similar to HSCs. Likewise, temporal RNAseq analysis of 3GF reprogrammed cells reveals a progression through an intermediate cell with a shared endothelial and hematopoietic gene profile, with acquisition of hematopoietic identity later in the reprogramming. Clusters of reprogrammed cells with a cobblestone-like appearance emerge after AFT024 co-culture that subsequently give rise to colonies consisting of various hematopoietic lineages. Limiting dilution analysis (LDA) of day 15 sorted GGF and 3GF cells on AFT024 revealed a significantly higher stem cell frequency when reprogramming with 3GF. 3GF cells reprogrammed to day 15 and sorted for CD49f display short-term multilineage engraftment potential upon intrahepatic transplant in newborn NSG mice. Taken together, these results demonstrate improvements in our human hemogenic induction strategy, bringing us one step closer to applying these findings to translational medicine.
Notlar:
School code: 1734
Tüzel Kişi Ek Girişi:
Mevcut:*
Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(682343.1) | 682343-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
On Order
Liste seç
Bunu varsayılan liste yap.
Öğeler başarıyla eklendi
Öğeler eklenirken hata oldu. Lütfen tekrar deneyiniz.
:
Select An Item
Data usage warning: You will receive one text message for each title you selected.
Standard text messaging rates apply.