Engraftment of engineered ES cell–derived cardiomyocytes but not BM cells restores contractile function to the infarcted myocardium

Abstract

Cellular cardiomyoplasty is an attractive option for the treatment of severe heart failure. It is, however, still unclear and controversial which is the most promising cell source. Therefore, we investigated and examined the fate and functional impact of bone marrow (BM) cells and embryonic stem cell (ES cell)–derived cardiomyocytes after transplantation into the infarcted mouse heart. This proved particularly challenging for the ES cells, as their enrichment into cardiomyocytes and their long-term engraftment and tumorigenicity are still poorly understood. We generated transgenic ES cells expressing puromycin resistance and enhanced green fluorescent protein cassettes under control of a cardiac-specific promoter. Puromycin selection resulted in a highly purified (>99%) cardiomyocyte population, and the yield of cardiomyocytes increased 6–10-fold because of induction of proliferation on purification. Long-term engraftment (4–5 months) was observed when co-transplanting selected ES cell–derived cardiomyocytes and fibroblasts into the injured heart of syngeneic mice, and no teratoma formation was found (n = 60). Although transplantation of ES cell–derived cardiomyocytes improved heart function, BM cells had no positive effects. Furthermore, no contribution of BM cells to cardiac, endothelial, or smooth muscle neogenesis was detected. Hence, our results demonstrate that ES-based cell therapy is a promising approach for the treatment of impaired myocardial function and provides better results than BM-derived cells.

Functional Expression and Regulation of Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels (HCN) in Mouse iPS Cell-derived Cardiomyocytes afterUTF1-Neo Selection

“…An alpha-myosin heavy chain ( αMHC )-promoter-driven puromycin resistance was introduced in order to purify miPS-CMs after differentiation [17]. The mES cell line D3-αPIG44, carrying a puromycin-resistance ( αMHC -promoter) and expressing eGFP [20], was used as control. To maintain their undifferentiated state, mESCs and miPSCs were grown on irradiated, mitotically inactive murine embryonic fibroblasts (MEFs) and cultured with 1 U/μl LIF (leukemia inhibitory factor, ESGRO, Millipore, Billerica, MA, USA).…”

Live monitoring of small vessels during development and disease using the flt-1 promoter element

“…ES cell differentiation was performed using embryoid body (EB) formation via mass culture [11] or hanging drop method [36]. For differentiation, cells were cultivated in Iscove's MEM high-glucose medium (Invitrogen, Karlsruhe, Germany) supplemented with 20% FCS plus additives in the absence of LIF.…”

Skeletal Extracellular Matrix Supports Cardiac Differentiation of Embryonic Stem Cells: a Potential Scaffold for Engineered Cardiac Tissue

“…The purified GFP-positive mESC-CMs were harvested by mass culture protocol as previously described [23]. Puromycin (10 µg/mL, InvivoGen) was added at day 9 and day 12 during the whole differentiation period [25]. Between day 14 and 16 of differentiation, the mESC-CMs were collected and digested into single cells by trypsin and used for following experiments.…”