Non-Enzymatic Generation of Placenta Single Cells From Third Trimester Human Placenta

Joy U Ameloko; Idowu A Aimola; Hanneda A Fomukong; Aliyu Salihu; Abdullahi B Salau; Zeenat B Kudan; Ruben S Baba; Solomon Dunason; Habiba H Abubakar; Kalen E Audu; Joy B Karma; Peculiar N Okoro; Abduljabar Musa; Musa Kana; Benedict Anchang

doi:10.1002/cpz1.70237

Non-Enzymatic Generation of Placenta Single Cells From Third Trimester Human Placenta

Curr Protoc. 2025 Oct;5(10):e70237. doi: 10.1002/cpz1.70237.

Authors

Joy U Ameloko^{1

2}, Idowu A Aimola^{1

2}, Hanneda A Fomukong^{1

2

3}, Aliyu Salihu^{1

2}, Abdullahi B Salau^{1

2}, Zeenat B Kudan^{1

2}, Ruben S Baba^{1

2}, Solomon Dunason⁴, Habiba H Abubakar⁴, Kalen E Audu¹, Joy B Karma^{1

2}, Peculiar N Okoro^{1

2}, Abduljabar Musa^{1

2}, Musa Kana⁴, Benedict Anchang⁵

Affiliations

¹ Africa Center of Excellence for Neglected Tropical Diseases and Forensic Biotechnology, Ahmadu Bello University, Zaria, Nigeria.
² Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria.
³ Department of Biochemistry, Capital City University Kano, Kano, Nigeria.
⁴ Kaduna Infant Development Birth Cohort Project, Kaduna, Nigeria.
⁵ Biostatistics and Computational Biology Branch, National Institute of Environmental Health Sciences, Durham, North Carolina.

PMID: 41129293
DOI: 10.1002/cpz1.70237

Abstract

The placenta is a heterogeneous and complex organ with multiple cell types, posing a challenge for the maternal-fetal medicine field to implement single-cell technologies for a deeper characterization of this essential organ. Several protocols use enzymes to digest the tissue and generate single-cell suspensions, but this approach has several shortcomings, including the loss and reduced viability of cells. In this article, we describe a non-enzymatic approach to generate single-cell suspensions from placental tissue with high yield and viability for single-cell RNA sequencing. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Preparation of single-cell suspension from freshly collected third trimester human placental sections Basic Protocol 2: Visualization and determination of cell viability Alternate Protocol: Counting cells using a hemocytometer.

Keywords: non‐enzymatic; placenta cell viability; single‐cell RNA sequencing; single‐cell genomics; third trimester placenta; transcriptomics.

MeSH terms

Cell Survival
Female
Humans
Placenta* / cytology
Pregnancy
Pregnancy Trimester, Third
Single-Cell Analysis* / methods