Comparison of different human cell derived induced pluripotent stem cells and embryonic stem cells in tuning embryoid body differentiation

doi:10.12307/2021.014

Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (25): 4019-4024.doi: 10.12307/2021.014

Comparison of different human cell derived induced pluripotent stem cells and embryonic stem cells in tuning embryoid body differentiation

Wei Yunjian1, 2, 3, Zhang Fengbo1, 2, 3, 4, Long Ping1, 2, 3, Jiang Xinxing1, 2, 3, Ma Yanlin1, 2, 3, Sun Fei1, 2, 3, 5, Li Qi1, 2, 3#br#

1Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; 2Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 570102, Hainan Province, China; 3Haikou Key Laboratory for Preservation of Human Genetic Resource, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; 4Department of Obstetrics, Anhui Province Maternity & Child Health Hospital, Hefei 230000, Anhui Province, China; 5Department of Obstetrics and Gynecology, NanFang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China

Received:2020-06-10 Revised:2020-06-16 Accepted:2020-07-31 Online:2021-09-08 Published:2021-03-27
Contact: Wei Yunjian, Research assistant, Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 570102, Hainan Province, China; Haikou Key Laboratory for Preservation of Human Genetic Resource, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China Zhang Fengbo, Master, Physician, Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 570102, Hainan Province, China; Haikou Key Laboratory for Preservation of Human Genetic Resource, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Department of Obstetrics, Anhui Province Maternity & Child Health Hospital, Hefei 230000, Anhui Province, China
About author:Sun Fei, MD, Physician, Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 570102, Hainan Province, China; Haikou Key Laboratory for Preservation of Human Genetic Resource, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Department of Obstetrics and Gynecology, NanFang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China Li Qi, Master, Researcher, Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China; Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou 570102, Hainan Province, China; Haikou Key Laboratory for Preservation of Human Genetic Resource, the First Affiliated Hospital of Hainan Medical University, Haikou 570102, Hainan Province, China
Supported by:
the Major Science and Technology Program of Hainan Province, No. ZDKJ2017007 (to MYL); Hainan Provincial Natural Science Foundation of China, No. 2019CXTD408 (to MYL); the National Natural Science Foundation of China, No. 81960283 (to MYL), No. 81660433 (to MYL), No. 81460034 (to LQ), No. 81460236 (to JXX)

Abstract

Abstract: BACKGROUND: Induced pluripotent stem cells (iPSCs) without virus vector integration can be successfully induced by small molecule compounds and plasmid transfection. Similar to embryonic stem cells, iPSCs can differentiate freely into hematopoietic progenitor cells. However, it is not clear whether exogenous factors and human cells from different sources affect the differentiation of iPSCs and hematopoietic progenitor cells.
OBJECTIVE: To compare the gene expression differences between iPSCs of different cell sources and embryoid bodies of embryonic stem cells, so as to explore the clinical transformation of iPSCs.
METHODS: The iPSCs and human embryonic stem cells were constructed from amniotic cells-derived cells and urine cells-derived cells. The different days of embryoid bodies were collected to test the expression of pluripotent genes Oct4, Sox2 and Nanog, endoderm marker gene GATA4, mesoderm marker gene MSX1, ectoderm marker gene PAX6, hematopoietic stem cell marker genes CD34/CD43.
RESULTS AND CONCLUSION: (1) The expression of Oct4/Sox2/Nanog in embryoid bodies from amniotic fluid cells derived iPSCs was almost undetectable at 8 days, which was different from urine cells derived iPSCs and embryoid bodies embryonic stem cells. They could sustain expression until 16 days. (2) The expression peak of GATA4/MSX1/PAX6 in embryoid bodies from amniotic fluid cells derived iPSCs was at 4 days, and sustained expression at 8-12 days. The expression peak of GATA4/MSX1/PAX6 in embryoid bodies from urine cells derived iPSCs and embryonic stem cells was at 4-8 days, and sustained expression at 12-16 days. The expression level of three layer-differentiation markers for embryoid bodies from urine cells derived iPSCs were significantly higher than iPSCs from amniotic fluid cells and embryonic stem cells. (3) At 12 days, the ratio of hematopoietic stem cell was the most in embryoid bodies from amniotic fluid cells and urine cells derived iPSCs. However, the ratio in the embryoid bodies from embryonic stem cells was at 4 days. The hematopoietic stem cell ratio was higher in embryoid bodies from urine cells derived iPSCs than that in amniotic fluid cells and embryonic stem cells. (4) Embryoid bodies from different cells derived iPSCs and embryonic stem cells all had pluripotency and the ability of three layer-differentiation, but they were distinctive. Urine cells derived iPSCs may replace embryonic stem cells as hematopoietic cell differentiation model in vitro.

Key words: stem cells, induced pluripotent stem cells, embryonic stem cells, embryoid bodies, pluripotency, hematopoietic differentiation, hematopoietic stem cells, amniotic fluid cells, urine cells

CLC Number:

Wei Yunjian, Zhang Fengbo, Long Ping, Jiang Xinxing, Ma Yanlin, Sun Fei, Li Qi. Comparison of different human cell derived induced pluripotent stem cells and embryonic stem cells in tuning embryoid body differentiation[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(25): 4019-4024.

Figures/Tables 4

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Comparison of different human cell derived induced pluripotent stem cells and embryonic stem cells in tuning embryoid body differentiation

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