Secondary sphere formation enhances the functionality of cardiac progenitor cells.
Loss of cardiomyocytes impairs cardiac function after myocardial infarction (MI). Recent studies suggest that cardiac stem/progenitor cells could repair the damaged heart. However, cardiac progenitor cells are difficult to maintain in terms of purity and multipotency when propagated in two-dimensional culture systems. Here, we investigated a new strategy that enhances potency and enriches progenitor cells. We applied the repeated sphere formation strategy (cardiac explant → primary cardiosphere (CS) formation → sphere-derived cells (SDCs) in adherent culture condition → secondary CS formation by three-dimensional culture). Cells in secondary CS showed higher differentiation potentials than SDCs. When transplanted into the infarcted myocardium, secondary CSs engrafted robustly, improved left ventricular (LV) dysfunction, and reduced infarct sizes more than SDCs did. In addition to the cardiovascular differentiation of transplanted secondary CSs, robust vascular endothelial growth factor (VEGF) synthesis and secretion enhanced neovascularization in the infarcted myocardium. Microarray pathway analysis and blocking experiments using E-selectin knock-out hearts, specific chemicals, and small interfering RNAs (siRNAs) for each pathway revealed that E-selectin was indispensable to sphere initiation and ERK/Sp1/VEGF autoparacrine loop was responsible for sphere maturation. These results provide a simple strategy for enhancing cellular potency for cardiac repair. Furthermore, this strategy may be implemented to other types of stem/progenitor cell-based therapy.
Cardiovascular Center & Department of Internal Medicine, Seoul National University, Seoul, Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Hyun-Jai
HJ
Cho
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea; Department of Biomedical Sciences, Seoul National University Graduate School, Seoul, Korea.
Ho-Jae
HJ
Lee
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Seock-Won
SW
Youn
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Seok-Jin
SJ
Koh
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Joo-Yun
JY
Won
Department of Biomedical Sciences, Seoul National University Graduate School, Seoul, Korea.
Yeon-Ju
YJ
Chung
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Hyun-Ju
HJ
Cho
Cardiovascular Center & Department of Internal Medicine, Seoul National University, Seoul, Korea.
Chang-Hwan
CH
Yoon
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Sae-Won
SW
Lee
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Eun Ju
EJ
Lee
Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Yoo-Wook
YW
Kwon
Cardiovascular Center & Department of Internal Medicine, Seoul National University, Seoul, Korea.
Hae-Young
HY
Lee
National Research Laboratory for Cell Physiology, Department of Physiology, Seoul National University College of Medicine, Seoul, Korea.
Sang Hun
SH
Lee
National Research Laboratory for Cell Physiology, Department of Physiology, Seoul National University College of Medicine, Seoul, Korea.
Won-Kyung
WK
Ho
Cardiovascular Center & Department of Internal Medicine, Seoul National University, Seoul, Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
Young-Bae
YB
Park
Cardiovascular Center & Department of Internal Medicine, Seoul National University, Seoul, Korea; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea; Department of Biomedical Sciences, Seoul National University Graduate School, Seoul, Korea; World Class University Program, Department of Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Korea. Electronic address: hyosoo@snu.ac.kr.
Hyo-Soo
HS
Kim
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D009203:Myocardial Infarction / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology; D018919:Neovascularization, Physiologic; D020411:Oligonucleotide Array Sequence Analysis; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D018487:Ventricular Dysfunction, Left / Q000378:metabolism / Q000473:pathology / Q000628:therapy*
D019040:E-Selectin; D007136:Immunoglobulins; D034741:RNA, Small Interfering; C090698:SP1 antigen; D042461:Vascular Endothelial Growth Factor A; D048049:Extracellular Signal-Regulated MAP Kinases