Analysis of Menstrual Blood Stromal Cells Reveals SOX15 Triggers Oocyte-Based Human Cell Reprogramming

Abstract

Summary Cell reprogramming has revolutionized cell and regenerative biology field. However, human iPS derivation remains inefficient and variable. A better knowledge of molecular processes and the rationale underlying the importance of somatic cell origin is crucial to uncover reprogramming mechanisms. Here, we analyze the molecular profile of different human somatic cell types. We show menstrual blood-derived stromal cells (MnSCs) have a distinct, reprogramming prone, profile, and we identify SOX15 from their oocyte-related signature as a prominent responsible candidate. SOX15 orchestrates an efficient oocyte-based reprogramming combination when overexpressed with the also oocyte-enriched histone chaperone ASF1A and OCT4 and, through specific mechanism, generates iPSCs with distinguishable pluripotent state that further present higher differentiation capacity than canonical iPSCs. Our work supports the presence of different pluripotency states in reprogramming and the importance of using metaphase-II oocyte and MnSCs information to provide alternative reprogramming combinations and, importantly, to improve and understand pluripotency acquisition.

Protocol to Reprogram Human Menstrual Blood-Derived Stromal Cells to Generate AOX15-iPSCs

“…Retroviral particles are prepared from pMX-backbone based vectors (from Cell Biolabs, find information in Key Resources Table ) encoding reprogramming factors ASF1A, SOX15 and OCT4 (AOX15)(please find pMX-ASF1A, pMX-SOX15 and pMX-OCT4 vector description in Lopez-Caraballo et al., 2020 )( Figure 3 ). Retroviral particles will be used to over express AOX15 combination in MnSCs.…”

“…We expect over 80% of the cells to be positive for mentioned pluripotency markers. Additionally, qRT-PCR can be performed to confirm the expression of pluripotency genes (including OCT4, SOX2, hTERT, DNM3TB, KLF4, NANOG, ZFP42, among others) ( Lopez-Caraballo et al., 2020 ).…”

“…Pluripotency is not a single state. Further characterization of the functional status of reprogrammed cells' pluripotency, including differentiation capacity, efficiency, and other molecular signatures can be performed ( Liu et al., 2017 ; Lopez-Caraballo et al., 2020 ; Mishra et al., 2018 ). To this end, a list of recommended antibodies for immunocytochemistry is provided in the Key Resources Table .…”

“…We describe an oocyte-based reprogramming combination to generate AOX15-iPSCs that can be used to study different states of pluripotency. For complete details on the use and execution of this protocol, please refer to Lopez-Caraballo et al. (2020) .…”

“…For complete details on the use and execution of this protocol, please refer to Lopez-Caraballo et al. (2020) .…”