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Induce Female Bone Marrow Stem Cells to Differentiate into Stra8~+ Cells and FSHR~+ Cells

Author: MiMeiLing
Tutor: ZouTing
School: Nanchang University
Course: Physiology
Keywords: Sertoli cells All-trans retinoic acid Female bone marrow stem cells FSHR Stra8
CLC: R329
Type: Master's thesis
Year: 2008
Downloads: 85
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Abstract

Objective: To explore the Sertoli cells (Sertoli cells, SCs) and female bone marrow stem cells (female bone marrow stem cells, FBMSCs) Isolation and culture techniques; observation SCs and all-trans retinoic acid (all-trans retinoic acid, ATRA) FBMSCs growth, proliferation; impact observed of SCs and of ATRA induced FBMSCs to the FSHR and Stra8 cell differentiation. Method: two-step enzymatic digestion and Tris-HCl hypotonic method separated 12-16d SD rat testis of SCs the whole bone marrow adherent law on separation of young female SD rats FBMSCs; co-cultured with the MTT colorimetric detection SCs and FBMSCs double room ( transwell membrane, the pore size of 0.4μm, allowing soluble molecules through) and ATRA culture FBMSCs survival and proliferation; for detection FBMSCs induced differentiation detected by RT-PCR germline stem cells into mitosis before meiosis-specific expression of the gene Stra8 The mRNA and ovarian granulosa cell specific expression of the gene FSHR mRNA expression induced differentiation of FBMSCs detection FBMSCs, SCs FSHR expression by immunocytochemistry, cell FSHR protein expression Western blot detection FBMSCs differentiation. Observed testicular SCs and ATRA induced adherent subculture the second generation FBMSCs to of Stra8 (Stra8 positive) and FSHR (FSHR-positive) cell differentiation. Results: 1 of SCs and FBMSCs were cultured group (co-culture group) and FBMSCs join ATRA culture group (ATRA group), co-culture group in cultured first 1 to 5 days proliferation was significantly greater than the control group (P lt; 0.01) , ATRA group to the fifth day of the first day of culture proliferation are greater than the control group, which on the third day, the fourth day there is a significant difference (P lt; 0.05). Six days to seven days, the number of cells in the control group slightly increased by co-culture group and ATRA the group FBMSCs cells decline is significantly reduced compared with the control group (P lt; 0.01), showed the early promotion of proliferation, the late increase in apoptosis biphasic effect. 2 the coculture group cultured for 7 days and ATRA group FBMSCs part differentiation become large and small, were round or spherical cells, differentiation of the large, round cells surrounded by many small cells surround. Control group FBMSCs cells are relatively uniform fusiform. 3 immunocytochemistry showed that co-culture group training 3,5,7 days FBMSCs partially differentiated medium-sized spherical or round FSHR-positive cells were clustered distribution, with the incubation time, the cell volume increases. RT-PCR showed the co-culture group and the ATRA group cultured for 7 days FBMSCs have Stra8mRNA of FSHR mRNA expression of the control group, no expression; western blot showed that co-culture and ATRA groups to cultivate the seven days FBMSCs have FSHR protein expression, and the control group. not expressed. Show that co-culture group and the ATRA group training FBMSCs could be induced to differentiate into FSHR and Stra8 cells. Conclusions: 1. Of SCs and FBMSCs were cultured group and join the ATRA group culture of FBMSCs growth, proliferation have pre-promote the proliferation of late caused a biphasic effect of the decline in the number of cells. 2. SCs FBMSCs co-culture group and ATRA group culture can be induced FBMSCs expression germline stem cells into mitosis before meiosis-specific expression of gene Stra8 mRNA that FBMSCs with germ cell differentiation of pluripotent stem cells. 3. SCs and FBMSCs training group and join the the ATRA group of culture can be induced FBMSCs expression in ovarian granulosa cell-specific expression of the gene FSHR mRNA and protein, FSHR-positive cells rendered as medium-sized spherical or round cells were clustered distribution, as the culture The prolonged cell volume increases, differentiation of pluripotent stem cells to the particle-like cells that FBMSCs having. 4. ATRA and SCs could in vitro induced FBMSCs differentiation Stra8 and FSHR cells, similar to the role of, speculated that ATRA SCs secrete soluble factor can induce FBMSCs of differentiation.

Induce Female Bone Marrow Stem Cells to Differentiate into Stra8~+ Cells and FSHR~+ Cells

Abstract

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