Geometric control of cardiomyogenic induction in human pluripotent stem cells.
نویسندگان
چکیده
Although it has been observed that aggregate size affects cardiac development, an incomplete understanding of the cellular mechanisms underlying human pluripotent stem cell-derived cardiomyogenesis has limited the development of robust defined-condition cardiac cell generation protocols. Our objective was thus to elucidate cellular and molecular mechanisms underlying the endogenous control of human embryonic stem cell (hESC) cardiac tissue development, and to test the hypothesis that hESC aggregate size influences extraembryonic endoderm (ExE) commitment and cardiac inductive properties. hESC aggregates were generated with 100, 1000, or 4000 cells per aggregate using microwells. The frequency of endoderm marker (FoxA2 and GATA6)-expressing cells decreased with increasing aggregate size during early differentiation. Cardiogenesis was maximized in aggregates initiated from 1000 cells, with frequencies of 0.49±0.06 cells exhibiting a cardiac progenitor phenotype (KDR(low)/C-KIT(neg)) on day 5 and 0.24±0.06 expressing cardiac Troponin T on day 16. A direct relationship between ExE and cardiac differentiation efficiency was established by forming aggregates with varying ratios of SOX7 (a transcription factor required for ExE development) overexpressing or knockdown hESCs to unmanipulated hESCs. We demonstrate, in a defined, serum-free cardiac induction system, that robust and efficient cardiac differentiation is a function of endogenous ExE cell concentration, a parameter that can be directly modulated by controlling hESC aggregate size.
منابع مشابه
Comparison of cardiomyogenic potential among human ESC and iPSC lines.
We recently reported that, following induction of clumps of pluripotent H1 human embryonic stem cells (hESCs) with activin-A and Bmp4 in defined medium for 5 days, widespread differentiation of rhythmically contracting cardiomyocytes occurs within 3-4 weeks. In this study, the same approach was used to assess whether human induced pluripotent stem cells (hiPSCs), which may theoretically provide...
متن کاملLarge-Scale Expansion of Human Embryonic and Induced Pluripotent Stem Cells for Cell Therapy Applications
Successful isolation, derivation and culturing of human pluripotent stem cells, including human embryonic stem cells (hESCs) and human induced pluripotent stem (hiPSCs) cells in laboratory scale has opened new horizones for cell therapy applications such as tissue engineering and regenerative medicine. However, most of the cell therapy protocols using these unique cells require large number of ...
متن کاملA Quick update from the Past to Current Status of Human Pluripotent Stem Cell-derived Hepatocyte culture systems
Pluripotent stem cells (PSCs) may be offered as an unlimited cell source for the hepatocyte generation. The generation of hepatocytes from stem cells in vitro would provide an alternative cell source for applications in drug discovery and cell transplantation. In this review, we discuss different approaches to generate pluripotent stem cell-derived hepatocytes, advantages, limitations for each ...
متن کاملP-50: Elongating and Elongated Spermatids Manufactured In Vitro from Non-Human Primate Pluripotent Stem Cells
Background: We have recently shown that human embryonic (hESCs) and induced pluripotent stem cells (hiPSCs) can differentiate into advanced spermatogenic cells including round spermatids by in vitro culture (Easley et al., Direct differentiation of human pluripotent stem cells into haploid spermatogenic cells. Cell Reports 2, 440-446 2012) and also, in collaboration, that rhesus spermatogonial ...
متن کاملThe Proliferation Study of hiPS Cell-Derived Neuronal Progenitors on Poly-Caprolactone Scaffold
Introduction: The native inability of nervous system to regenerate, encourage researchers to consider neural tissue engineering as a potential treatment for spinal cord injuries. Considering the suitable characteristics of induced pluripotent stem cells (iPSCs) for tissue regeneration applications, in this study we investigated the adhesion, viability and proliferation of neural progenitors (de...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Tissue engineering. Part A
دوره 17 15-16 شماره
صفحات -
تاریخ انتشار 2011