A New Xeno-free Medium for Human Mesenchymal Stem Cell Expansion ex vivo

Lee, Don-Ching; Wang, Mei-Chih; Yu, Wei-Lin; Wu, Jiun-Yi; Shiu, Heng-Cheng; Cheung, Chi-Shiu; Tsai, Wen-Che

doi:10.30491/jabr.2024.449555.1708

A New Xeno-free Medium for Human Mesenchymal Stem Cell Expansion ex vivo

Document Type : Original Article

Authors

Don-Ching Lee ¹

Mei-Chih Wang ²^{, 3}

Wei-Lin Yu ³

Jiun-Yi Wu ⁴

Heng-Cheng Shiu ⁵

Chi-Shiu Cheung ¹

Wen-Che Tsai ¹

¹ GeneDireX, Inc., Taoyuan, Taiwan

² Department of Biotechnology and Pharmaceutical Technology, Yuanpei University of Medical Technology, Hsinchu, Taiwan

³ Biomedical Technology & Device Research Laboratories, Industrial Technology Research Institute, Hsinchu, Taiwan

⁴ Department of Orthopaedic Surgery, Hsinchu branch, National Taiwan University Hospital, Hsinchu, Taiwan

⁵ Department of Gynecology and Obstetrics, National Taiwan University Hospital, Taipei, Taiwan

10.30491/jabr.2024.449555.1708

Abstract

Introduction: An optimal culture medium that can rapidly and efficiently proliferate cells ex vivo is very crucial for developing mesenchymal stem cells (MSCs)-based tissue engineering and regenerative medicine. We developed a set of MSCs ex vivo proliferation medium, mscGO^TM XF, which consists of a basal medium and a screened human platelet lysate.
Materials and Methods: In this study, the developed mscGO^TM XF medium was prepared, and then testified by human MSCs isolated from bone marrow, umbilical cord, and fat tissue. The proliferation, surface markers, differentiation, and chromosomal stability of MSCs cultured in mscGO^TM XF medium were investigated.
Results: The mscGO^TM XF medium could sustain MSCs at a high proliferation rate, with the population doubling time of 16 to 39 hours (depending on the type and passage number of MSCs). The proliferated MSCs could express CD105, CD90, and CD73, lack expression of CD34 and CD45; and maintain the capacity to differentiate into adipocytes, osteoblasts, and chondrocytes. Additionally, G-Band karyotyping data confirmed chromosome stability in the duration of cell culture at passage 5 and passage 7.
Conclusions: The mscGOTM XF medium could sustain MSCs proliferation ex vivo and exhibit the potential to be developed into a clinical-grade cell culture medium kit.

Keywords

Xeno-free Medium

Mesenchymal Stem Cells

Cell Proliferation

Cell Differentiation