Effects of Human Umbilical Cord Mesenchymal Stem Cells on the Proliferation and Differentiation of Endometrium in Rats with Intrauterine Adhesions

doi:10.12280/gjfckx.20201077

Abstract

Abstract:

Objective: To evaluate the effect of human umbilical cord mesenchymal stem cells (hUCMSC) on the proliferation and differentiation of injured endometrium in rats with intrauterine adhesions. Methods: 20 SD rats of intrauterine adhesions model was established on the principle of self-own control. After 14 days,they were randomly divided into the stem cell group (n=10) and the control group (n=10). hUCMSC were isolated from human umbilical cord specimens, and then cultured and identified for the further study. hUCMSC in passage 3 were used to transplant into the stem cell group of uterus with intrauterine adhesions and phosphate buffer saline solution were injected into the control group of uterus with intrauterine adhesions. Immunofluorescence staining was used to detect the distribution of human nuclear antibodies, endometrial epithelial cell marker cytokeratin (CK) antibody and mesenchymal cell marker vimentin antibody in rat uterus to evaluate the location and differentiation of hUCMSC in rat uterus. Immunohistochemical staining was used to observe the expression of proliferating cell nuclear antigens Ki-67 and CK in hUCMSC and mRNA of Ki-67 and CK in rat uterus were detected by quantitative reverse transcription polymerase chain reaction. Results: The modeling side of endometrial tissue of the rat uterine showed narrow of the uterine cavity,lack of the endometrial glands and luminal epithelium,increased areas without cell structure in the stroma. Cultured cells were conformed to have the characteristics of hUCMSC in terms of cell morphology and immunophenotype. In the stem cell group, red and green fluorescence were co-developed in the same cell in the co-stained sections of human nucleus and vimentin. The average optical density and mRNA expression of Ki-67 and CK in the stem cell group were higher than those in the control group, and the differences were statistically significant (P<0.05). Conclusions: hUCMSC can survive and colonize in the injured endometrium and may improve the tissue repair microenvironment by up-regulating the expression of Ki-67 and CK, rather than by differentiation into endometrial cells.

Key words: Umbilical cord, Mesenchymal stormal cells, Uterus, Tissue adhesions, Uterine adhesion, Cell proliferation, Ki-67 antigen

WANG Sha, GUO Zheng-chen, TANG Yi-qun, DUAN Hua. Effects of Human Umbilical Cord Mesenchymal Stem Cells on the Proliferation and Differentiation of Endometrium in Rats with Intrauterine Adhesions[J]. Journal of International Obstetrics and Gynecology, 2021, 48(3): 309-313.

Figures/Tables 6

References 17

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试剂	使用量
2×TS Reaction Mix	10 μL
Anchored Oligo（dT）18（0.5 μg/μL）	1 μL
TransStart?RT/RI Enzyme Mix	1 μL
RNA	1 μg
RNase free water	补充体系至20 μL

试剂	使用量
2×TS Reaction Mix	10 μL
Anchored Oligo（dT）18（0.5 μg/μL）	1 μL
TransStart?RT/RI Enzyme Mix	1 μL
RNA	1 μg
RNase free water	补充体系至20 μL

名称	引物	序列（5′→3′）	片段长度（bp）
GAPDH	正向	TACCCACGGCAAGTTCAACG	92
	反向	CACCAGCATCACCCCATTTG
Ki-67	正向	AAGACGAAGATCAGGCAGGA	200
	反向	CAACAATCAGGTTGGCTTCA
CK	正向	TCCTCAGCCATGTCTTCCTATG	287
	反向	CTAAAACTTCCACCGCGTCCT

名称	引物	序列（5′→3′）	片段长度（bp）
GAPDH	正向	TACCCACGGCAAGTTCAACG	92
	反向	CACCAGCATCACCCCATTTG
Ki-67	正向	AAGACGAAGATCAGGCAGGA	200
	反向	CAACAATCAGGTTGGCTTCA
CK	正向	TCCTCAGCCATGTCTTCCTATG	287
	反向	CTAAAACTTCCACCGCGTCCT

组别	n	Ki-67		CK
组别	n	平均光密度	mRNA	平均光密度	mRNA
干细胞组	10	0.34±0.07	0.64±0.19	0.34±0.08	0.37±0.12
对照组	10	0.16±0.06	0.22±0.17	0.17±0.07	0.17±0.08
t		6.17	5.21	5.06	4.385
P		<0.01	<0.01	<0.01	<0.01