ReproFF2 Customer Feedback

The purpose of feeder-free culture with ReproFF2

Below are the examples of the purposes to conduct feeder-free culture with ReproFF2.

Contamination of the feeder cells can be eliminated by adopting feeder-free culture, which makes it possible to conduct the researches which have been difficult or impossible with the conventional feeder-dependent culture system.

  • For extraction of DNA/RNA without contamination of those from the feeder cells
  • For regenarative medicine-directed researches
  • For removal of anti-differentiation effects by the feeder cells
  • For promoting viral infection

Culture results with ReproFF2 by the customers

Below are the tables summarizing the culture results of human ES/iPS cells with ReproFF2 by the customers. ReproFF2 has been successfully adopted by many laboratories for various human ES/iPS cell lines.

Use of ReproFF2 for human ES cell lines
Organization human ES cell lines Coating of the culture surface Culture result
Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University KhES-1 Laminin-5 Y
Research Institute National Center for Global Health and Medicine KhES-3 Matrigel Y
Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine, IMSUT KhES-3 Matrigel Y
Department of Physiology, School of Medicine, Keio University H9 Matrigel Y
Use of ReproFF2 for human iPS cell lines
>Organization human iPS cell line Method for establishing the iPS cell line Coating of the culture surface Culture result
Institute of Molecular Embryology and Genetics, Kumamoto University 201B7 HDF/Retrovirus Matrigel Y
Research Institute National Center for Global Health and Medicine SeV-iPS HUVEC/Sendai virus Matrigel Proliferation of feeder-like cells was also observed
RIKEN Center for Developmental Bilogy Line 1 CellStart Y
Line 2 Laminin-5 Y
(Proliferation of feeder-like cells was also observed when Cell Start was used)
Line 3 Laminin-5 Y
(The attachment of the cells was not excellent, but the undifferentiated state could be maintained even with the cell line which was difficult to keep undifferentiated in feeder-dependent culture.)
(Proliferation of feeder-like cells was also observed when Cell Start was used)
Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine, IMSUT TkDA 3-4 HDF/4 retrovirus Matrigel Y
Department of Physiology, School of Medicine, Keio University 201B7 HDF/HDF/4 retrovirus Matrigel Y

Comments from the customers

Dr. Nobuaki Shiraki, Assistant Professor
Institute of Molecular Embryology and Genetics, Kumamoto University

Cell line: 201B7

“It was very useful that we did not require feeder cells and the medium needed changing only on alternate days. The cells remained viable through freezing and thawing. Endoderm differentiation was similar in cells cultured with ReproStem/MEF and with ReproFF2/Matrigel.”

RIKEN Center for Developmental Bilogy

Cell line: several iPS cell lines established in house

“Some cell lines that are difficult to maintain in an undifferentiated state in feeder-dependent culture could be maintained using ReproFF2.

The condition of the culture was exacerbated when the cells were transferred from feeder-dependent culture to feeder-free culture with ReproFF2 , but it recovered after several passages.

There are some tips including ensuring that the size of the colonies is suitable for passaging, but the procedure itself is straightforward.”

Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine, IMSUT

Cell line: KhES-3、TkDA3-4 (estalished from HDF with retroviral transduction)

“Some trial and error was necessary to switch from feeder-dependent culture to feeder-free culture with ReproFF2, but finally, the culture was successful.”

Department of Physiology, School of Medicine, Keio University

Cell line: H9, 201B7, etc.

“It was easy to switch from feeder-dependent culture to feeder-free culture with ReproFF2 for any cell line.”

Research Institute National Center for Global Health and Medicine

Cell line: KhES-3, SeV-iPS (established from HUVEC with Sendai viral transduction of the 4 factors)

“KhES3 was cultured without any difficulty in feeder-free culture with ReproFF2 after switching from feeder-dependent culture.

In the SeV-iPS cell line, we observed apparently differentiated cells proliferating around the colonies. However, the iPS cell colonies were intact, and immunostaining showed that the colonies were positive for Oct3/4.”