Migratory and anti-fibrotic programmes define the regenerative potential of human cardiac progenitors

DOI: 10.1038/s41556-022-00899-8

Authors: Christine M. Poch; Kylie S. Foo; Maria Teresa De Angelis; Karin Jennbacken; Gianluca Santamaria; Andrea Bähr; Qing-Dong Wang; Franziska Reiter; Nadja Hornaschewitz; Dorota Zawada; Tarik Bozoglu; Ilaria My; Anna Meier; Tatjana Dorn; Simon Hege; Miia L. Lehtinen; Yat Long Tsoi; Daniel Hovdal; Johan Hyllner; Sascha Schwarz; Stefanie Sudhop; Victoria Jurisch; Marcella Sini; Mick D. Fellows; Matthew Cummings; Jonathan Clarke; Ricardo Baptista; Elif Eroglu; Eckhard Wolf; Nikolai Klymiuk; Kun Lu; Roland Tomasi; Andreas Dendorfer; Marco Gaspari; Elvira Parrotta; Giovanni Cuda; Markus Krane; Daniel Sinnecker; Petra Hoppmann; Christian Kupatt; Regina Fritsche-Danielson; Alessandra Moretti; Kenneth R. Chien; Karl-Ludwig Laugwitz

Step 1: iPSC/ESC maintainance

day 0 ~ 1

culture “adherent” human ESC in “E8” for “1” days. “ESC” should be “unspecified regulation” for “NKX2-5”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 2: Primitive streak 1

day 1 ~ 2

culture “adherent” human ESC in “RPMI” for “1” days. “ESC” should be “unspecified regulation” for “NKX2-5”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “1” days.

Step 3: mesoderm 1

day 2 ~ 4

culture “adherent” human ESC in “RPMI” for “2” days. “ESC” should be “unspecified regulation” for “NKX2-5”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.

Ideally, “mesoderm” should express “EOMES”, “MESP1”, “LGR5” at the end of culture.

Step 4: mesoderm 2

day 4 ~ 6

culture “adherent” human ESC in “RPMI” for “2” days. “ESC” should be “unspecified regulation” for “NKX2-5”. For ECM, use “Matrigel” at “N/A” “N/A”. Medium should contain factors shown in the following table, and should be changed once in “2” days.