Miniaturized droplet microarray platform enables maintenance of human induced pluripotent stem cell pluripotency

Yanxi Liu; Shraddha Chakraborty; Chatrawee Direksilp; Johannes M Scheiger; Anna A Popova; Pavel A Levkin

doi:10.1016/j.mtbio.2021.100153

Miniaturized droplet microarray platform enables maintenance of human induced pluripotent stem cell pluripotency

Mater Today Bio. 2021 Oct 25:12:100153. doi: 10.1016/j.mtbio.2021.100153. eCollection 2021 Sep.

Authors

Yanxi Liu¹, Shraddha Chakraborty¹, Chatrawee Direksilp², Johannes M Scheiger^{1

3}, Anna A Popova¹, Pavel A Levkin^{1

4}

Affiliations

¹ Institute of Biological and Chemical Systems - Functional Molecular Systems (IBCS-FMS), Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, Eggenstein-Leopoldshafen 76344, Germany.
² The Petroleum and Petrochemical College (PPC), Chulalongkorn University, Soi Chulalongkorn 12, Phayathai road, Pathumwan, 10330 Bangkok, Thailand.
³ Institute of Chemical Technology and Polymer Chemistry (ITCP), Karlsruhe Institute of Technology (KIT), Engesserstraße 20, 76131 Karlsruhe, Germany.
⁴ Institute of Organic Chemistry (IOC), Karlsruhe Institute of Technology (KIT), Kaiserstraße 12, 76131 Karlsruhe, Germany.

Abstract

The capacity of human induced pluripotent stem cells (hiPSCs) for indefinite self-renewal warrants their application in disease modeling, drug discovery, toxicity assays and efficacy screening. However, their poor proliferation ability, inability to adhere to surfaces without Matrigel coating and tendency to spontaneously differentiate in vitro hinder the application of hiPSCs in these fields. Here we study the ability to culture hiPSCs inside 200 nL droplets on the droplet microarray (DMA) platform. We demonstrate that (1) hiPSCs can attach to the Matrigel (MG)-free surface of DMA and show good viability after 24 h culture; (2) hiPSC do not spontaneously differentiate when cultured on the MG-free surface of DMAs; (3) culturing of hiPSCs in 200 nL as compared to 2 mL culture leads to higher expression of the Nanog pluripotency marker. Overall, the results demonstrate the possibility to culture undifferentiated hiPSCs in 200 nL droplets on DMA, thereby opening the possibility for high-throughput screenings of hiPSCs with various factors without compromising the results through the involvement of animal-derived materials, such as Matrigel.

Keywords: Cell culture substrates; Droplet microarray; High-throughput screening; Human induced pluripotent stem cells; Stem cell pluripotency; Xeno-free culture.